Macrocyclic compounds useful as inhibitors of kinases and HSP90

ABSTRACT

Disclosed are macrocyclic compounds of formulae I-V, which are analogs of the pochonin resorcylic acid lactones, and processes for the preparation of the compounds. The compounds disclosed are useful as inhibitors of kinases and Heat Shock Protein 90 (HSP 90). Also disclosed are pharmaceutical compositions comprising an effective kinase-inhibiting amount or an effective HSP90-inhibiting amount of the compounds and methods for the treatment of disorders that are mediated by kinases and HSP90.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationNos. 60/837,154, filed Aug. 11, 2006, and 60/858,731, filed Nov. 13,2006.

FIELD OF THE INVENTION

The present invention is directed to novel derivatives, analogs andintermediates of the natural products radicicol and the pochonins, andto their syntheses. The present invention is further directed to use ofthese compounds as inhibitors of kinases and of the enzyme family knownas heat shock protein 90 (HSP90).

BACKGROUND OF THE INVENTION

In the mid-1950's, it was discovered that phosphorylation can reversiblyalter the function of enzymes by means of protein kinases which catalyzephosphorylation, or by protein phosphatases which are involved in thedephosphorylation step. These reactions play an essential role inregulating many cellular processes, especially signaling transductionpathways. In the late 1970's, the Rous sarcoma virus (v-Src)'stransforming factor was discovered to be a protein kinase, and alsotumor-promoting phorbol esters were found to be potent activators ofprotein kinase C, revealing the first known connection between diseaseand abnormal protein phosphorylation. Since then transductionmechanistic defects have been found to cause numerous oncogenicprocesses and to have a role in diabetes, inflammatory disorders, andcardiovascular diseases. (T. Hunter, Cell, 100:113-127 (2000); P. Cohen,Nat. Rev. Drug Discov., 1:309 (2002)). Thus selective kinase andphosphatase inhibitors have emerged as important drug targets, andinhibition of kinase phosphorylation activity is one of the mostpromising strategies for chemotherapy. Three kinase inhibitor drugs arealready approved: Gleevec, which inhibits Abl, and Iressa and Tarceva,which both inhibit EGFR.

Gleevec (imatinib) - STI 571 inhibitor of the Abelson kinase; approvedfor chronic myelogenous leukemia

Tarceva (erlotinib) - OSI774 R1 = CCH; R2 = H; R3 = R4 = CH₂OCH₃ Iressa(gefitinib) - ZD1839 R1 = Cl; R2 = F; R3 = H; R4 =

inhibitors of epidermal growth factor receptor (EGFR); approved for lungcancer

Modulation of protein activity by kinase-mediated phosphorylation orphosphatase-mediated dephosphorylation of a serine, threonine ortyrosine residue is at the center of most signal transductionmechanisms. (T. Hunter, Cell, 100:113 (2000)). Small molecule inhibitorssuch as 6-dimethylaminopurine and staurosporine were instrumental inelucidating the importance of such phosphorylation mechanisms and shedlight on the biological function of kinases. Kinases bind to ATP with aK_(m) of 0.1-10 μM, and transfer the γ-phosphate group selectively to aspecific residue of a given protein. The core domain of kinases,consisting of the ATP-binding site with the residues involved inphosphotransfer reaction, are highly conserved throughout the kinome.(G. Manning et al., Science, 298:1912 (2002)). This led to thespeculation that inhibitors targeting this highly conserved ATP-bindingpocket would not only have to compete with ATP present at highconcentration (mM) but would necessarily lack selectivity. The discoverythat modified purines such as (R)-roscovitine were potent and fairlyselective inhibitors (L. Meijer and E. Raymond, Acc. Chem. Res., 36:417(2003)) refuted that notion and inspired the synthesis of combinatoriallibraries around the purine scaffold (Y. T. Chang et al., Chem. Biol.,6:361 (1999); S. Ding et al., J. Am. Chem. Soc., 124:1594 (2002)),yielding important leads. (N. S. Gray et al., Science, 281:533 (1998);M. Knockaert et al., Chem. Biol., 7:411 (2000)).

Macrocyclic resorcylic acid lactones have also been investigated in thisrespect. The archetypes of this class of compounds are radicicol and therelated pochonins, which are a structurally related group of secondarymetabolites isolated from cultures of the clavicipitaceous hyphomycetePochonia genus, such as Pochonia chlamydosporia var. catenulate strainP0297. See, e.g., V. Hellwig et al., J. Natural Prod., 66(6):829-837(2003). Halohydrin and oxime derivatives of radicicol were prepared andevaluated for their v-src tyrosine kinase inhibitory, antiproliferative,and antitumor in vitro activity (T. Agatsuma et al., Bioorg. & Med.Chem., 10(11):3445-3454 (2002).

Like kinases, heat shock proteins (HSPs) interact with ATP and areimportant targets for controlling disease, however they have a differentmechanistic effect. Immediately after exposure to a stress such as heat,hypoxia or acidosis, cells in most tissues rapidly escalate productionrate of the HSPs. It is now believed that heat HSPs are molecularchaperones, i.e., they prevent improper associations and assist in thecorrect folding of other cellular proteins collectively termed clientsand substrates. HSP's are also found in association with tumors andother pathophysiological conditions. In fact, chaperone proteinsfacilitate the survival of tumor cells in stressful environments byfacilitating tolerance of alterations inside the cell. HSPs areubiquitous, highly conserved among the species, and usually classifiedby molecular weight to the following major families: HSP100, HSP90,HSP70, HSP60 and small HSPs. These families have structural andfunctional differences, but work cooperatively at different stages ofprotein folding. HSP90 has attracted particular attention due to itsassociation with many types of signaling molecules such as v-Src and Rafthat play a critical role in malignant transformation and metastasisdevelopment. Thus, HSP90 inhibitors are desired for designingchemotherapies, and also for elucidating the interplay in complexsignaling networks.

Heat Shock Protein 90's (Hsp90s) are ubiquitous chaperone proteins thatmaintain the proper conformation of many “client” proteins (see Kamalet. al. Trends Mol. Med. 2004, 10, 283-290; Dymock et. al. Expert Opin.Ther. Patents 2004, 14, 837-847; Isaacs et. al. Cancer Cell, 2003, 3,213; Maloney et. al. Expert Opin. Biol. Ther. 2002, 2, 3-24 and Richteret. al. J. Cell. Physiol. 2001, 188, 281-290), and are involved infolding, activation and assembly of a wide range of proteins, includingkey proteins involved in signal transduction, cell cycle control andtranscriptional regulation. Researchers have reported that HSP90chaperone proteins are associated with important signaling proteins,such as steroid hormone receptors and protein kinases, including, e.g.,Raf-1, EGFR, v-Src family kinases, Cdk4, and ErbB-2 (Buchner, TIBS,1999, 24, 136-141; Stepanova et. al., Genes Dev. 1996, 10, 1491-502; Daiet. al., J. Biol. Chem. 1996, 271, 22030-4). Studies further indicatethat certain co-chaperones, e.g., Hsp70, p60/Hop/Sti1, Hip, Bag1,HSP40/Hdj2/Hsj1, immunophilins, p23, and p50, may assist HSP90 in itsfunction (see for example Caplan, Trends in Cell Biol., 1999, 9,262-268). Inhibition of Hsp90 causes these client proteins to adoptaberrant conformations, and these abnormally folded proteins are rapidlyeliminated by the cell via ubiquitinylation and proteasome degradation.Interestingly, the list of Hsp90 client proteins includes a series ofnotorious oncogenes. Four of them are clinically validated cancertargets: HER-2/neu (Herceptin® (trastuzumab)), Bcr-Abl (Gleevec®(imatinib mesylate)), the estrogen receptor (tamoxifen), and theandrogen receptor (Casodex® (bicalutamide)), while the others play acritical role in the development of cancer. Some of the most sensitiveHsp90 clients are involved in growth signalling (Raf-1, Akt, cdk4, Src,Bcr-Abl, etc). In contrast, few tumor suppressor genes, if any, seem tobe clients of Hsp90 (for lists of client proteins see Pratt et. al. Exp.Biol. Med. 2003, 228, 111-133; Workman et. al. Cancer Lett. 2004, 206,149-157 and Zhang et. al. J. Mol. Med. 2004, 82, 488-499.), andconsequently, inhibition of Hsp90 has an overall anti-proliferativeeffect. In addition, some client proteins are involved in otherfundamental processes of tumorigenesis, namely apoptosis evasion (e.g.Apaf-1, RIP, Akt), immortality (e.g. hTert), angiogenesis (e.g. VEGFR,Flt-3, FAK, HIF-1), and metastasis (c-Met).

However medicinal HSP inhibitors must be selective because HSPs alsoplay a constructive role. Under non-stressed conditions, HSP90 is one ofthe most abundant proteins present in the eukaryotic cells, representingbetween 1-2% of the total cellular protein content and increasing onlyabout two-fold when cells are stressed. Upon binding with the nativeclient HSP90 is an essential housekeeper, e.g., for folding of nascentpolypeptides, transporting proteins across membranes, and for normalprotein turnover. Moreover, HSP90 plays a crucial role inpost-translational regulation of signaling molecules, leading to theiractivation. HSP90 rarely functions alone but instead works withchaperone HSP70, with co-chaperones (HSP40, CDC37/p50, AHAL, p23), andwith accessory proteins.

The numerous client proteins of HSP90 play a crucial role in growthcontrol, cell survival and development processes, and those clients areknown to include receptor tyrosine kinases, serine/threonine kinases,steroid hormone receptors, transcription factors and telomerase.Oncogenic mutants of clients are also clients themselves but have higherrequirements for HSP90 function, for instance the mutant v-SRC tyrosinekinase requires more protein-folding capability from HSP90's cooperativeassembly of proteins (Y. Xu et al., Proc. Natl. Acad. Sci. U.S.A.,96:109 (1999); H. Oppermann et al., Ibid., 78:1067 (1981); L. Whitesellet al., Ibid., 91:8324 (1994). Likewise, mutations of thetumor-suppressor protein p53 lead to the most common molecular geneticdefect found in human cancers, and most p53 mutants show extendedinteractions with HSP90 (probably because of aberrant conformations),preventing their usual ubiquitylation and subsequent degradation by theproteasome (M. V. Blagoskionny et al., Ibid., 93:8379 (1996). Howeverdespite its ubiquitous participation, HSP90's clients are mostlypro-growth signaling proteins, and its chaperoning function is subvertedduring oncogenesis, leading to development of malignant transformationand the maintenance of transformed phenotypes.

In addition to anti-cancer and antitumorgenic activity, HSP90 inhibitorshave also been implicated in a wide variety of other utilities,including use as anti-inflammation agents, anti-infectious diseaseagents, agents for treating autoimmunity, agents for treating ischemia,and agents useful in treating neurodegenerative diseases and inpromoting nerve regeneration (see M. Waza et al, Nature Med. 11:1088(2005); Rosen et al., WO 02/09696; PCT/US01/23640; Degranco et al., WO99/51223; PCT/US99/07242; Gold, U.S. Pat. No. 6,210,974 B1). There arereports in the literature that fibrogenetic disorders including but notlimited to scleroderma, polymyositis, systemic lupus, rheumatoidarthritis, liver cirrhosis, keloid formation, interstitial nephritis,and pulmonary fibrosis may be treatable. (Strehlow, WO 02/02123;PCT/US01/20578).

Ansamycins and other HSP90 inhibitors thus hold great promise for thetreatment and/or prevention of many types of disorders. However, many ofthe natural-product derived Hsp90 inhibitors exhibit pharmaceuticaldeficiencies; their relative insolubility makes them difficult toformulate and administer, and they are not easily synthesized andcurrently must, at least in part, be generated through fermentation.Further, the dose limiting toxicity of ansamyins is hepatic. Forexample, the semi-synthetic inhibitor17-allylamino,17-desmethoxy-geldanamycin (17-AAG), currently in phase IIclinical trials, is expensive to manufacture, difficult to formulate(the NCI clinical protocol consists of injecting a DMSO solution of17-AAG) and at present administered only parenterally. Although the17-dimethylaminoethylamino analog (17-DMAG) is more soluble, it exhibitsall of the side effects of 17-AAG as well as gastrointestinalhemorrhaging in preclinical toxicity studies (Glaze et. al. Proc. Am.Assoc. Cancer. Res. 2003, 44, 162-162 and Eiseman et. al. CancerChemother. Pharmacol. 2005, 55, 21-32). Radicicol (RC), another naturalproduct Hsp90 inhibitor, is poorly water-soluble and is inactive intumor xenograft models. Semi-synthetic oxime derivatives of radicicolprovide better solubility and substantially improved the pharmacologicalprofile in murine models, but are still limited to intravenousadministration (Ikuina et. al. J. Med. Chem. 2003, 46, 2534-2541.Furthermore, radicicol and its oximes contain an oxirane ring which hasbeen viewed as a liability for stability and toxicity, prompting thesynthesis of cycloproparadicicol: Yang et. al. J. Am. Chem. Soc. 2004,126, 7881 and 2003, 125, 9602-9603.) Despite the potential ofansamycins, alternative HSP90 inhibitors are therefore needed.

Fully synthetic, orally active inhibitors of Hsp90 have been sought inorder to provide more flexible dosing schedule options, and to possiblyavoid the side-effects of the natural product inhibitors. Chiosis et al.described the design and synthesis of purine analogs that mimicgeldanamycin and other ansamycins in their ability to bind the ATPbinding pocket of, and thus inhibit, HSP90. See International PatentApplication PCT/US01/46303 (WO 02/36075; Chemistry & Biology 8:289-299(2001). The specific compounds that Chiosis et al. described included atrimethoxybenzyl entity substituted at positions 3, 4, and 5. Usinggel-binding assays, these were shown to bind HSP90 approximately 20-foldless avidly than 17-AAG.

More recently, other novel non-natural product Hsp90 inhibitors havebeen reported (e.g. PU3 and CCT018159; see Chiosis et. al. Bioorg. Med.Chem. Lett. 2002, 10, 3555-3564; Vilenchik et. al. Chem. Biol. 2004, 11,787-797; Chiosis et. al. WO 0236075, 2002; Drysdale et. al. WO 03/055860A1, 2003; Wright et. al. Chem. Biol. 2004, 11, 775-785; Dymock et. al.Bioorg. Med. Chem. Lett. 2004, 14, 325-328; Dymock et. al. J. Med. Chem.2005, 48, 4212-4215. Structure of Hsp90 in complex with PU3 pdb code1UY6, and with PU24FCl: pdb code 1UYF and Clevenger et. al. Org. Lett.2004, 6, 4459-4462). The structures of these inhibitors were designedusing the crystal structures of Hsp90 in complex with ATP, geldanamycin,or radicicol. The 8-benzyladenines such as PU3 were designed to adoptthe same C-shaped conformation as geldanamycin (Chiosis et. al. CurrentCancer Drug Targets, 2003, 3, 371-376) with the adenine ring pointing tothe adenine-binding site (hinge region), and the trimethoxybenzene ringemulating the H-bond accepting nature of the quinone ring ofgeldanamycin. (The benzene ring of PU3 was not designed to have exactlythe same orientation as the quinone ring of geldanamycin. Rather, thetrimethoxybenzene moiety was designed to point in the same generaldirection and form a hydrogen bond with Lys112, an amino acid whichforms a hydrogen bond with the quinone ring of geldanamycin.) Therecently obtained crystal structure of Hsp90 in complex with PU3confirmed that the purine ring occupies the position normally occupiedby ADP/ATP, but the benzene ring points in a direction opposite to thepredicted one, to form a r-stacking interaction with Phe138.Nevertheless, PU3 inhibits Hsp90 (HER-2 degradation assay, HER-2 IC₅₀=40μM) and afforded a valuable starting point for further optimization.Structure-activity studies based on PU3 led to the more active PU24FCl(HER-2 IC₅₀=1.7 μM) which was subsequently also co-crystallized withHsp90. When PU24FCl was formulated in DMSO/EtOH/phosphate-bufferedsaline 1:1:1 and administered intraperitoneally to mice bearing MCF-7xenograft tumors, it induced at 100-300 mg/kg down-regulation of HER-2and Raf-1, a pharmacodynamic response consistent with Hsp90 inhibition,and at 200 mg/kg it significantly repressed tumor growth. Very highdoses (500-1000 mg/kg) of PU24FCl were required to observe a similarpharmacodynamic response upon oral administration, and no8-benzyladenine has been reported to inhibit tumor growth by the oralroute. In our hands, PU24FCl proved to be too insoluble to beeffectively formulated and delivered orally. So far, despite extensiveSAR studies to improve potency and pharmaceutics properties, Hsp90inhibitors have not demonstrated activity in animal models of humancancer (xenografts) when administered orally.

The discovery of the 8-benzyladenines led to the design of8-sulfanyladenines (Kasibhatla et. al. WO 3037860, 2003 and Llauger et.al. J. Med. Chem. 2005, 48, 2892-2905), exemplified by8-(2-iodo-5-methoxy-phenylsulfanyl)-9-pent-4-ynyl-9H-purin-6-ylamine,which exhibited excellent potency in several cell-based assays, but waspoorly soluble in water and did not have sufficient oral bioavailabilityin clinically acceptable formulations.

When HSP90 is inhibited, its clients are degraded, i.e., the unfoldedprotein is ubiquitinated, followed by proteasome-mediated hydrolysis.Most of the inhibitors reported so far bind to the N-terminal domain(vide infra), but some are reported to interact with the C-terminaldomain; HSP90 has binding sites for ATP in both locations. The functionof HSP90's C terminus is not entirely clear, but compounds interactingin this domain clearly impair HSP90 function and have anti-cancereffects. Some resorcylic acid lactones have been found to inhibit HSP90,thus natural products radicicol and geldanamycin (P. Delmotte and J.Delmotte-Plaquee, Nature (London), 171:344 (1953); and C. DeBoer et al.,J Antibiot (Tokyo), 23:442 (1970), respectively) were shown to suppressthe transformed phenotype of cell expressing activated Src (H. J. Kwonet al., Cancer Research, 52:6926 (1992); Y. Uehara et al., Virology,164:294 (1988)). Related compounds such as herbimycin have been reportedto have similar effects (S. Omura et al., J Antibiot (Tokyo), 32:255(1979).

Other resorcylic acid lactones (RALs) studied in this respect include17-allylamino-17-demethoxygeldanamycin (17AAG) (D. B. Solit et al.,Clin. Cancer Res., 8:986 (2002); L. R. Kelland et al., J. Natl. CancerInst., 91:1940 (1999)); 17DMAG (J. L. Eiseman et al., Cancer Chemother.Pharmacol., 55:21-32 (2005)); IPI-504 (J. Ge et al., J. Med. Chem.,49:4606 (2006); oxime derivatives such as KF25706 (S. Soga, et al.,Cancer Res., 59:2931 (1999)) and KF55823 (S. Soga et al., CancerChemotherapy and Pharmacology, 48:435 (2001)); and Danishefsky et al.'scycloproparadicicol (A. Rivkin et al., Ibid., 44:2838 (2005)).Structurally related variants include chimeric inhibitors havingradicicol's carboxyresorcinol and the geldanamycin's benzoquinone (R. C.Clevenger and B. S. Blagg, Org. Lett., 6:4459 (2004); G. Shen and B. S.Blagg, Ibid., 7:2157 (2004); G. Shen et al., J. Org. Chem., 71:7618(2006)).

Radicicol-Based HSP90 Inhibitors

Chimeric Inhibitors of HSP90

Purines such as PU3 have been studied in an effort to design smallmolecules that fit HSP90's ATP binding site (G. Chiosis, et al., ChemBiol 8, 289-299 (2001); G. Chiosis, et al., Bioorg. Med. Chem., 10:3555(2002); L. LLauger, et al., J. Med. Chem. 48:2892 (2005); H. He et al.,Ibid., 49:381 (2006); M. A. Biamonte et al., Ibid., 49:817 (2006)).

Purine-Based Designed HSP90 Inhibitors

Pyrazoles (1-35) (M. G. Rowlands et al., Anal. Biochem., 327:176 (2004);B. W. Dymock et al., J. Med. Chem., 48:4212 (2005)) andbenzothiazolothio-purines (1-36) (L. Zhang. et al., J. Med. Chem.,49:5352 (2006) have been reported recently also as small-moleculeinhibitors of these enzymes.

Other Classes of HSP90 Inhibitors

Radicicol has been of particular interest. A 14-member macrolide, andalso known as monorden, radicicol is a potent, highly competitive andhighly selective ligand for HSP90's ATP-binding pocket. HSP90 is anATPase rather than a kinase, and its ATP-binding pocket has a Bergeratfold (A. Bergerat et al., Nature, 386:414 (1997); R. Dutta and M.Inouye, Trends Biochem. Sci., 25:24 (2000)) which is distinct fromkinases' ATP-binding pockets. (S. M. Roe et al., J. Med. Chem., 42:260(1999)). Considerable interest in radicicol's medicinal applicationshave followed the initial findings. (See U.S. Pat. No. 6,946,456; andU.S. Patent Application Publication Nos. 2003-0211469, 2004-0102458,2005-0074457, 2005-0261263, 2005-0267087, 2006-0073151, 2006-0251574,2006-0269618, 2007-0004674, and 2007-0010432).

radicicol HSP-90 inhibitor (20 nM) cocrystal structure

pochonin C Helicase-primase inhibitor

LL-783,277 MEK inhibitor (4 nM)

hypothemycin Inhibits Ras function (cellular assay)

LL-Z1640-2: R = H; TAK-1 inhibitor (8.1 nM) F87-2509.04: R = OMe;Promotes degradation of ARE-containing genes (cellular assay)

aigialomycin D CDK1/cyclin B (5.7 mM) CDK5/p25 (5.8 mM)

Strikingly, some resorcylic macrolides that are close analogs ofradicicol are known to inhibit kinases but not HSP90. Indeed, LL-Z1640-2was found to be a potent and selective inhibitor of TAK1 kinase forwhich radicicol and other resorcylides were not active. (J.Ninomiya-Tsuji et al., J. Biol. Chem., 278:18485 (2003); P. Rawlins etal., Int. J. Immunopharma., 21:799 (1999); K. Takehana et al., Biochem.Biophys. Res. Comm., 257:19 (1999); A. Zhao et al., J. Antibiotics,52:1086 (1999)). Closely related LL-783,227, where one of the olefinshas been reduced, is a potent inhibitor of MEK kinase. (A. Zhao et al.,J. Antibiotics 52:1086 (1999)). Compound F87-2509.04 was found to inducedegradation of mRNA containing AU-rich elements (ARE) (T. Kastelic etal., Cytokine, 8:751 (1996)) and hypothemycin was found to inhibit theRas-mediated cellular signaling. (H. Tanaka et al., Jap. J. Cancer Res.,90:1139 (1999)). We have recently shown that aigialomycin D is a CDKinhibitor. (S. Barluenga et al., Angew. Chem., Int. Ed., 46(24):3951(2006)).

Other close analogs of radicicol do inhibit HSP90. Pochonin D is apotent inhibitor of HSP90. (E. Moulin et al., J. Am. Chem. Soc.,127(19):6999 (2005)). And pochonin A has been reported to be a 90 nMinhibitor of HSP90. Pochonin C was found to be an inhibitor of herpes'helicase-primase, which is an ATPase rather than a kinase. (V. Hellwiget al., J. Nat. Prod., 66:829 (2003)). Although radicicol and pochonin Care structurally very similar, they have very different conformations insolution, and different biological activities. (S. Barluenga et al.,Chem. Eur. J., 11:4935 (2005). Thus it appears the “floppiness” of themacrocyclic may play an essential role in inhibitory differences amongresorcylic acid macrolides, and in any case makes those effectsdifficult to predict by theoretical methods.

Some resorcylic acid macrolides had been known as kinase or phosphataseinhibitors (U.S. Pat. Nos. 5,674,892; 5,728,726; 5,731,343; and5,795,910), or to inhibit other enzymes (U.S. Pat. No. 5,710,174inhibiting FXIIIa catalysis of fibrin cross-linking). Resorcylic acidmacrolides were also employed for other medical indications (U.S. Pat.Nos. 3,453,367; 3,965,275; 4,035,504; 4,670,249; 4,778,821; 4,902,711;and 6,635,671).

Radicicol and the pochonins are natural products; intermediates forsynthesizing some of their analogues of them may be obtained byfermentation, however relying only upon those natural products or theirfermentation derivatives severely limits the range of compounds. Thus anumber of novel resorcylic acid macrolides have been synthesized. Manyof these are zearalane and related compounds in which the macrocyclicring contains no carbon-carbon double bond other than between carbons ofthe phenyl ring. (U.S. Pat. Nos. 3,373,038; 3,586,701; 3,621,036;3,631,179; 3,687,982; 3,704,249; 3,751,431; 3,764,614; 3,810,918;3,836,544; 3,852,307; 3,860,616; 3,901,921; 3,901,922; 3,903,115;3,957,825; 4,042,602; 4,751,239; 4,849,447; and 2005-0256183). Syntheseshave also been reported for resorcylic acid macrolides characterized byone double bond between ring carbons outside the phenyl ring. (U.S. Pat.Nos. 3,196,019; 3,551,454; 3,758,511; 3,887,583; 3,925,423; 3,954,805;and 4,088,658). Most of those are 14-member macrocycles, but syntheseshave also been reported for the 12-member macrocycle analogs. (U.S. Pat.Nos. 5,710,174; 6,617,348; and 2004-0063778. and PCT publication no. WO02/48135)

Syntheses have also been reported for radicicol-related compounds havingtwo non-aromatic double bonds and either a halide or a 1,2-oxo group(i.e., an epoxide) on the macrocyclic ring. (U.S. Pat. Nos. 4,228,079;5,597,846; 5,650,430; 5,977,165; 7,115,651; and Japanese patent documentnos. JP 6-279279A, JP 6-298764A, JP 9-202781A, JP 10-265381A2; and JP2000-236984). Syntheses of oximes of radicicol-related compounds aredisclosed in U.S. Pat. Nos. 5,977,165; 6,239,168; 6,316,491; 6,635,662;2001-0027208; 2004-0053990; Japanese patent document no. JP2003-113183A2; and PCT publication no. WO 99/55689 Synthesis ofcyclopropa-analogs of radicicol is disclosed in U.S. Pat. No. 7,115,651and PCT Publication No. WO 05/061481. Syntheses of some other resorcylicacid macrolide analogs are disclosed in U.S. patent publication no.2006-0247448 and in PCT publication no. WO 02/48135. Radicicol as wellas Pochonins A and C have also been synthesized. (S. Barluenga et al.,Angew. Chemie, 43(26):3467-3470 (2004); S. Barluenga et al., Chemistry—AEuropean Journal, 11(17):4935-4952 (Aug. 19, 2005); E. Moulin et al., etal., Organic Letters, 7(25):5637-5639 (Dec. 8, 2005).

Despite the progress described above, chemical biologists continue tosuffer from a limited ability to knock out specific kinase activity inorder to deconvolute the role of specific kinases within complexsignaling networks. Small molecules that can permeate cells have promisefor solving this problem. And it has become increasingly apparent thatthe biological function of kinases is often regulated by theirconformation, which is in turn dictated by their phosphorylation leveland by intra- and inter-molecular associations. Small moleculeinhibitors also have the potential to discriminate between differentconformations of a given kinase, thus small molecules offer a means todissect the respective functions of those conformation. Unfortunatelythe portfolio of known kinase inhibitors cannot yet support the fullrange of work to be done in parsing the roles of the various members ofthe kinome. This is not a merely academic pursuit, because therationality of drug design will continue to suffer until kinasemechanisms and their selectivity is understood.

Thus there is an ongoing need for kinase inhibitors and HSP90 inhibitorsthat have improved potency and selectivity. Moreover, the design andsynthesis of such inhibitors and of targeted libraries of inhibitorsremains challenging, thus there is an ongoing need for improvedsynthetic methods.

SUMMARY OF THE INVENTION

Novel analogs of the pochonin macrolides of formulae I, II, III, IV andV, tautomers thereof, pharmaceutically acceptable salts, solvates,esters or prodrugs thereof, and pharmaceutical compositions comprisingthe compounds for the treatment of kinase-mediated or HSP90-mediateddisorders are provided. Also presented are methods for the treatment ofkinase-mediated or HSP90-mediated disorders using the compounds offormulae I, II, III, IV and V. The compounds of the invention are activeas kinase inhibitors and inhibitors of HSP90. In addition, improvedprocesses for the preparation of the compounds that are amenable toautomated synthesis techniques are provided.

In a first principal embodiment of the invention, a compound of formulaI, a tautomer thereof, or a pharmaceutically acceptable salt, solvate,ester or prodrug thereof, is provided:

wherein:

-   -   R¹, R², R³, R⁴ and R⁵ are independently hydrogen, halogen,        nitro, cyano, alkyl, substituted alkyl, lower alkyl, alkenyl,        alkynyl, alkylaryl, aralkyl, aryl, heteroalkyl, alkylheteroaryl,        heterocyclyl, heteroaryl, OH, OR, NH₂, N(R)₂, SR, S(O)R, S(O)₂R,        —SO₂N(R)₂, —N(R)SO₂R, —N(CO)R, —N(CO)N(R)₂, —N(CO)OR, —O(CO)R,        —(CO)R, —(CO)OR, —(CO)N(R)₂, —O(CO)OR, or —O(CO)N(R)₂, wherein        each R can be the same or different;    -   A₁ and A₂ together are —CH₂—CH₂—, —CH═CH—, —CH(OH)—CH(OH)—,        —CH(OH)—CH(halogen)-, —CH(halogen)-CH(OH)—, 1,2-cyclopropadiyl,        or 1,2-oxirane;    -   B₁ and B₂ together are —CH₂—CH₂— or B₁ and B₂ together represent        a covalent bond, wherein each R can be the same or different;    -   X¹ is hydrogen, halogen, OH, OR, NH₂, N(R)₂, NH—OR, SR, S(O)R,        S(O)₂R, —NH—O—(CH₂)_(n)—CO₂—R, —NH—O—(CH₂)_(n)—CON(R)₂; or X₁        together with X₂ or X₃ represents a covalent bond, wherein each        R can be the same or different;    -   X⁴ and X⁵ together are ═O, ═S, ═N—OR, ═N—O—(CH₂)_(n)COOR,        ═N—O—(CH₂)_(n)CON(R)₂, ═N—N(R)₂, ═N—N—SOR or ═N—N—SO₂R, wherein        the groups —OR, —O—(CH₂)_(n)COOR, —O—(CH₂)_(n)CON(R)₂, —N(R)₂,        —N—SOR or —N—SO₂R bound to the nitrogen may be in Z- or        E-configuration; or one of X₄ and X₅ is hydrogen and the other        is OH, OR, O(CO)R, O(CO)OR, O(CO)N(R)₂, —(CH₂)_(n)C(O)OR, or        —(CH₂)_(n)C(O)N(R)₂, and n is 0, 1, 2, or 3; or one of X⁴ and X⁵        together with X⁶ represents a covalent bond and the other of X⁴        and X⁵ is OH, OR, O(CO)R, O(CO)OR, —N(R)₂ or O(CO)N(R)₂, wherein        each R can be the same or different;    -   X⁶ is hydrogen or X⁶ together with one of X⁴ and X⁵ represents a        covalent bond; and    -   R is hydrogen, alkyl, substituted alkyl, lower alkyl, acyl,        aryl, alkaryl, arylalkyl including benzyl, heteroalkyl,        heteroaryl, heterocyclyl, a protecting group; or two R on the        same nitrogen are taken together with the nitrogen to form a 5-8        membered heterocyclic or heteroaryl ring; and    -   n is 0, 1, 2 or 3.

In a second embodiment of the invention, a compound of formula II, atautomer thereof, or a pharmaceutically acceptable salt, solvate, esteror prodrug thereof, is provided:

-   -   wherein, R, R¹, R², R³, R⁴, R⁵, A₁, A₂, X¹, X² and X³ are as        defined for formula I above; and    -   R⁷ is ═O, ═S, ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂,        ═N—N(R)₂, ═N—N—SOR or ═N—N—SO₂R, wherein the groups —OR,        —O—(CH₂)_(n)COOR, —O—(CH₂)_(n)CON(R)₂, —N(R)₂, —N—SOR or —N—SO₂R        bound to the nitrogen may be in Z- or E-configuration; and        wherein R can be the same or different.

In one subembodiment of formula II, A₁ and A₂ together are —CH═CH—.

In another subembodiment of formula II, A₁ and A₂ together are—CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or —CH(halogen)-CH(OH)—.

In still another subembodiment of formula II, A₁ and A₂ together are1,2-oxirane.

In third embodiment of the invention, a compound of formula III, atautomer thereof, or a pharmaceutically acceptable salt, solvate, esteror prodrug thereof, is provided:

-   -   wherein, R¹, R², R³, R⁴, R⁵, A₁, A₂, X¹, X² and X³ are as        defined for formula I above;    -   and R is hydrogen, alkyl, arylalkyl, acyl or a protecting group.

In one subembodiment of formula III, A₁ and A₂ together are —CH═CH—.

In another subembodiment of formula III, A₁ and A₂ together are—CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or —CH(halogen)-CH(OH)—.

In still another subembodiment of formula III, A₁ and A₂ together are1,2-oxirane.

In a fourth embodiment of the present invention, a compound of formulaIV, a tautomer thereof, or a pharmaceutically acceptable salt, solvate,ester or prodrug thereof, is provided:

-   -   wherein, R, R¹, R², R³, R⁴, R⁵, A₁ and A₂ are as defined for        formula I above; and R⁶ is hydrogen, OR, or N(R)₂.

In one subembodiment of formula IV, A₁ and A₂ together are —CH═CH—.

In another subembodiment of formula IV, A₁ and A₂ together are—CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or —CH(halogen)-CH(OH)—.

In a further subembodiment of formula IV, A₁ and A₂ together are1,2-oxirane.

In a fifth embodiment of the invention, a compound of formula V, atautomer thereof, or a pharmaceutically acceptable salt, solvate, esteror prodrug thereof, is provided:

-   -   wherein, R, R¹, R², R³, R⁴, R⁵, A₁ and A₂, are as defined for        formula I above; R⁶ is (CH₂)_(n)C(O)OR, or —(CH₂)_(n)C(O)N(R)₂;        and    -   n is 0, 1, 2 or 3.

In one subembodiment of formula V, A₁ and A₂ together are —CH═CH—.

In another subembodiment of formula V, A₁ and A₂ together are—CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or —CH(halogen)-CH(OH)—.

In a third subembodiment of formula V, A₁ and A₂ together are1,2-oxirane.

In another embodiment, a pharmaceutical composition comprising aneffective kinase-inhibiting amount of a compound of formula I, II, III,IV or V in combination with a pharmaceutically acceptable carrier isprovided.

In another embodiment, a pharmaceutical composition comprising aneffective HSP 90-inhibiting amount of a compound of formula I, II, III,IV or V in combination with a pharmaceutically acceptable carrier isprovided. In some embodiments, the carrier is suitable for oral,parenteral, inhalation, topical or intradermal administration.

In still other embodiments, the pharmaceutical compositions comprisingthe compounds of formula I, II, III, IV or V comprises particles thatare less than about 2 microns average particle size. In still otherembodiments, the composition is incorporated into a biodegradable ornon-biodegradable polymer.

In one embodiment, the composition comprises an additive selected froman anti-oxidant, a buffer, a bacteriostat, a liquid carrier, a solute, asuspending agent, a thickening agent, a flavoring agent, a gelatin,glycerin, a binder, a lubricant, an inert diluent, a preservative, asurface active agent, a dispersing agent, a biodegradable polymer, orany combination thereof.

In another embodiment, the invention provides a method of treating apatient with a disease comprising administering to the patient with thedisease an effective amount of a compound of formula I, II, III, IV orV, wherein the disease is an autoimmune disease, an inflammatorydisease, a neurological or neurodegenerative disease, cancer, acardiovascular disease, allergy, asthma, or a hormone-related disease.

In one embodiment, a method of treating a patient with cancer isprovided comprising administering to the patient having the cancer aneffective cancer-treating amount of a compound of formula I, II, III, IVor V, wherein the cancer may be a solid tumor, a blood borne tumor,breast cancer, cancer of the ovary, cancer of the cervix, prostatecancer, cancer of the testis, cancer of the urinary tract, cancer of theesophagus, cancer of the larynx, glioblastoma, stomach cancer, skincancer, keratoacanthoma, lung cancer, epidermoid carcinoma, large cellcarcinoma, small cell carcinoma, lung adenocarcinoma, bone cancer, coloncancer, adenoma, cancer of the pancreas, adenocarcinoma, thyroid cancer,follicular carcinoma, undifferentiated carcinoma, papillary carcinoma,seminoma, melanoma, sarcoma, bladder carcinoma, liver carcinoma andbiliary passages, kidney carcinoma, myeloid disorders, lymphoiddisorders, Hodgkin's, hairy cells, buccal cavity cancer, pharynx cancer,lip cancer, tongue cancer, mouth cancer, cancer of the pharynx, cancerof the small intestine, colon-rectum cancer, cancer of the largeintestine, cancer of the rectum, brain cancer and cancer of the centralnervous system, or leukemia.

In another embodiment, the invention provides a method of treating apatient with a disease associated with undesirable neovascularizationcomprising administering to the patient with the undersirableneovascularization an effective amount of a compound of formula I, II,III, IV or V.

The disease associated with undesirable neovasculariation comprisesocular neovascular disease, diabetic retinopathy, retinopathy ofprematurity, corneal graft rejection, neovascular glaucoma andretrolental fibroplasias, epidemic keratoconjunctivitis, Vitamin Adeficiency, contact lens overwear, atopic keratitis, superior limbickeratitis, pterygium keratitis sicca, Sjögren's syndrome, acne rosacea,phylectenulosis, syphilis, Mycobacteria infections, lipid degeneration,chemical burns, bacterial ulcers, fungal ulcers, Herpes simplexinfections, Herpes zoster infections, protozoan infections, Kaposi'ssarcoma, Mooren's ulcer, Terrien's marginal degeneration, marginalkeratolysis, trauma, rheumatoid arthritis, systemic lupus,polyarteritis, Wegener's sarcoidosis, Scleritis, Steven-Johnson disease,pemphigoid, radial keratotomy, or corneal graph rejection, sickle cellanemia, sarcoid, pseudoxanthoma elasticum, Paget's disease, veinocclusion, artery occlusion, carotid obstructive disease, chronicuveitis/vitritis, Lyme's disease, systemic lupus erythematosis, Eales'disease, Bechet's disease, infections causing a retinitis orchoroiditis, presumed ocular histoplasmosis, Best's disease, myopia,optic pits, Stargart's disease, pars planitis, chronic retinaldetachment, hyperviscosity syndromes, toxoplasmosis, or post-lasercomplications.

In still another embodiment, a method of treating a patient with aninflammatory disease is provided comprising administering to the patientwith the inflammatory disease an effective amount of a compound offormula I, II, III, IV or V.

The inflammatory disease may be excessive or abnormal stimulation ofendothelial cells, atherosclerosis, vascular malfunctions, abnormalwound healing, inflammatory and immune disorders, Bechet's disease, goutor gouty arthritis, abnormal angiogenesis accompanying rheumatoidarthritis, skin diseases, psoriasis, diabetic retinopathy, retinopathyof prematurity, retrolental fibroplasia, macular degeneration, cornealgraft rejection, neovascular glaucoma or Osler Weber syndrome.

DETAILED DESCRIPTION OF THE INVENTION

Provided are novel compounds based on the resorcylic acid lactones thatare useful as inhibitors of kinases and HSP90. Also provided arecompositions comprising the compounds and processes for the preparationof the compounds. Use of the compounds for the inhibition of kinases andHSP-90, and a method for the treatment of kinase-mediated orHSP90-mediated diseases comprising administering an effectivekinase-inhibiting amount or an effective HSP90-inhibiting amount of acompound of formula I, II, III, IV or V to a patient with akinase-mediated or HSP90-mediated disease, are provided.

Compounds

In a first embodiment of the invention, a compound of formula I, atautomer thereof, or a pharmaceutically acceptable salt, solvate, esteror prodrug thereof, is provided:

-   -   wherein:    -   R¹, R², R³, R⁴ and R⁵ are independently hydrogen, halogen,        nitro, cyano, alkyl, substituted alkyl, lower alkyl, alkenyl,        alkynyl, alkylaryl, aralkyl, aryl, heteroalkyl, alkylheteroaryl,        heterocyclyl, heteroaryl, OH, OR, NH₂, N(R)₂, SR, S(O)R, S(O)₂R,        —SO₂N(R)₂, —N(R)SO₂R, —N(CO)R, —N(CO)N(R)₂, —N(CO)OR, —O(CO)R,        —(CO)R, —(CO)OR, —(CO)N(R)₂, —O(CO)OR, or —O(CO)N(R)₂, wherein        each R can be the same or different;    -   A₁ and A₂ together are —CH₂—CH₂—, —CH═CH—, —CH(OH)—CH(OH)—,        —CH(OH)—CH(halogen)-, —CH(halogen)-CH(OH)—, 1,2-cyclopropadiyl,        or 1,2-oxirane;    -   B₁ and B₂ together are —CH₂—CH₂— or B₁ and B₂ together represent        a covalent bond, wherein each R can be the same or different;    -   X¹ is hydrogen, halogen, OH, OR, NH₂, N(R)₂, NH—OR, SR, S(O)R,        S(O)₂R, —NH—O—(CH₂)_(n)—CO₂—R, —NH—O—(CH₂)_(n)—CON(R)₂; or X₁        together with X₂ or X₃ represents a covalent bond, wherein each        R can be the same or different;    -   X² and X³ are both hydrogen, or one of X₂ and X₃ is hydrogen and        the other together with X₁ represents a covalent bond;    -   X⁴ and X⁵ together are ═O, ═S, ═N—OR, ═N—O—(CH₂)_(n)COOR,        ═N—O—(CH₂)_(n)CON(R)₂, ═N—N(R)₂, ═N—N—SOR or ═N—N—SO₂R, wherein        the groups —OR, —O—(CH₂)_(n)COOR, —O—(CH₂)_(n)CON(R)₂, —N(R)₂,        —N—SOR or —N—SO₂R bound to the nitrogen may be in Z- or        E-configuration; or one of X₄ and X₅ is hydrogen and the other        is OH, OR, O(CO)R, O(CO)OR, O(CO)N(R)₂, —(CH₂)_(n)C(O)OR, or        —(CH₂)_(n)C(O)N(R)₂, and n is 0, 1, 2, or 3; or one of X₄ and X⁵        together with X⁶ represents a covalent bond and the other of X⁴        and X⁵ is OH, OR, O(CO)R, O(CO)OR, —N(R)₂ or O(CO)N(R)₂, wherein        each R can be the same or different;    -   X⁶ is hydrogen or X⁶ together with one of X⁴ and X⁵ represents a        covalent bond; and    -   R is hydrogen, alkyl, lower alkyl, acyl, aryl, alkaryl,        arylalkyl including benzyl, heteroalkyl, heteroaryl,        heterocyclyl, a protecting group; or two R on the same nitrogen        are taken together with the nitrogen to form a 5-8 membered        heterocyclic or heteroaryl ring; and    -   n is 0, 1, 2 or 3.

In one subembodiment of compound I, R¹ is H, halogen or heterocyclyl.

In another subembodiment of compound I, R⁵ is hydrogen, alkyl, loweralkyl, aryl, heteroaryl or arylalkyl.

In another subembodiment of compound I, A₁ and A₂ together are —CH₂—CH₂—or —CH═CH—.

In still another subembodiment of compound I, A₁ and A₂ together are—CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or CH(halogen)-CH(OH)—.

In a further subembodiment of compound I, A₁ and A₂ together are1,2-cyclopropadiyl, or 1,2-oxirane.

In second embodiment of the invention, a compound of formula II, atautomer thereof, or a pharmaceutically acceptable salt, solvate, esteror prodrug thereof, is provided:

wherein:

-   -   R, R¹, R², R³, R⁴, R⁵, X¹, X², X³, A₁ and A₂ are as defined        above for formula I; and    -   R⁷ is ═O, ═S, ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂,        ═N—N(R)₂, ═N—N—SOR or ═N—N—SO₂R, wherein the groups —OR,        —O—(CH₂)_(n)COOR, —O—(CH₂)_(n)CON(R)₂, —N(R)₂, —N—SOR or —N—SO₂R        bound to the nitrogen may be in a Z- or E-configuration; and        wherein R can be the same or different.

In one subembodiment of compound II, R¹ is H, halogen or heterocyclyl.

In another subembodiment of compound II, R⁵ is hydrogen, alkyl, loweralkyl, aryl, heteroaryl or arylalkyl.

In another subembodiment of compound II, A₁ and A₂ together are—CH₂—CH₂— or —CH═CH—.

In still another subembodiment of compound II, A₁ and A₂ together are—CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or —CH(halogen)-CH(OH)—.

In a further subembodiment of compound II, A₁ and A₂ together are1,2-cyclopropadiyl, or 1,2-oxirane.

In another subembodiment of compound II:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are —CH═CH—;    -   X¹ together with X² represent a covalent bond; and    -   R⁷ is ═O.

In another subembodiment of compound II:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are —CH═CH—;    -   X¹ together with X² represent a covalent bond; and    -   R⁷ is ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂,        ═N—N(R)₂, or ═N—N—SOR, wherein the groups —OR, —O—(CH₂)_(n)COOR,        —O—(CH₂)_(n)CON(R)₂, —N(R)₂, —N—SOR or —N—SO₂R bound to the        nitrogen may be in Z- or E-configuration.

In another subembodiment of compound II:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are —CH═CH— or —C(OH)—C(OH)—;    -   X¹ is hydrogen, halogen or NH—OR; and    -   R⁷ is ═O.

In another subembodiment of compound II:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are —CH═CH— or —C(OH)—C(OH)—;    -   X¹ is hydrogen, halogen or NH—OR; and    -   R⁷ is ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂, ═N—N(R)₂        or ═N—N—SOR.

In another subembodiment of compound II:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are —CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or        —CH(halogen)-CH(OH)—;    -   X¹ together with X² represent a covalent bond; and    -   R⁷ is ═O.

In another subembodiment of compound II:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are —CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or        —CH(halogen)-CH(OH)—;    -   X¹ together with X² represent a covalent bond; and    -   R⁷ is ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂,        ═N—N(R)₂, ═N—N—SOR, ═N—N—SO₂R.

In another subembodiment of compound II:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl or aryl;    -   A₁ and A₂ together are —CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or        —CH(halogen)-CH(OH)—;    -   X¹ is hydrogen, halogen, NH—O—(CH₂)_(n)COOR, or        NH—O—(CH₂)_(n)CON(R)₂ NH—OR; and    -   R⁷ is ═O, ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂,        ═N—N(R)₂, ═N—N—SOR, or ═N—N—SO₂R.

In still another subembodiment of compound II:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are 1,2-oxirane or 1,2-cyclopropadiyl;    -   X¹ together with X² represent a covalent bond; and    -   R⁷ is ═O, ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂,        ═N—N(R)₂, ═N—N—SOR, or ═N—N—SO₂R.

In still another subembodiment of compound II:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are 1,2-oxirane or 1,2-cyclopropadiyl;    -   X¹ is hydrogen, halogen, NH—OR, NH—O—(CH₂)_(n)COOR, or        NH—O—(CH₂)_(n)CON(R)₂; and    -   R⁷ is ═O, ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂,        ═N—N(R)₂, ═N—N—SOR, or ═N—N—SO₂R.

In still a further subembodiment of compound II:

-   -   R¹ is H or Cl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen;    -   A₁ and A₂ together —CH═CH—;    -   X¹ together with X² represent a covalent bond; and    -   R⁷ is ═O.

In still a further subembodiment of compound II:

-   -   R¹ is H or Cl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen;    -   A₁ and A₂ together —CH═CH—;    -   X¹ together with X² represent a covalent bond; and    -   R⁷ is ═N—OR, ═N—O—(CH₂)_(n)COOR, or ═N—O—(CH₂)_(n)CON(R)₂.

In yet another subembodiment of compound II:

-   -   R¹ is H or Cl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen;    -   A₁ and A₂ together are 1,2-oxirane;    -   X¹ together with X² represent a covalent bond; and    -   R⁷ is ═O.

In yet another subembodiment of compound II:

-   -   R¹ is H or Cl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen;    -   A₁ and A₂ together are 1,2-oxirane;    -   X¹ together with X² represent a covalent bond; and    -   R⁷ is ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂.

In a further subembodiment of formula II:

-   -   R¹ is H or Cl;    -   R² and R⁴ are OH;    -   R⁵ is hydrogen;    -   A₁ and A₂ together are —CH═CH—;    -   X¹ together with X² represent a covalent bond;    -   R⁷ is ═N—O—CH₂COOR or ═N—O—CH₂CON(R)₂; and    -   R is H, lower alkyl, or two R together on the same nitrogen form        a six-membered heterocyclyl ring.

In another embodiment of the invention, a compound of formula III, atautomer thereof, or a pharmaceutically acceptable salt, solvate, esteror prodrug thereof is provided:

wherein:

-   -   R, R¹, R², R³, R⁴, R⁵, X¹, X², X³, A₁ and A₂ are as defined        above for formula I.

In one subembodiment of compound III, X¹ and X² together form a doublebond.

In one subembodiment of compound III, R is hydrogen or acyl.

In one subembodiment of compound III, R¹ is H, halogen or heterocyclyl.

In another subembodiment of compound III, R₅ is hydrogen, alkyl, loweralkyl, aryl, heteroaryl or arylalkyl.

In another subembodiment of compound III, A₁ and A₂ together are—CH₂—CH₂— or —CH═CH—.

In still another subembodiment of compound III, A₁ and A₂ together are—CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or CH(halogen)-CH(OH)—.

In a further subembodiment of compound III, A₁ and A₂ together are1,2-cyclopropadiyl, or 1,2-oxirane.

In one subembodiment of formula III, a compound is provided wherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are —CH═CH—; and    -   X¹ together with X² represent a covalent bond.

In another subembodiment of formula III, a compound is provided wherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are —CH═CH—; and    -   X¹ is hydrogen, halogen, NH—OR, NH—O—(CH₂)_(n)COOR, or        NH—O—(CH₂)_(n)CON(R)₂.

In yet another subembodiment of formula III, a compound is providedwherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are —CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or        —CH(halogen)-CH(OH)—; and    -   X¹ together with X² represent a covalent bond.

In a further subembodiment of formula III, a compound is providedwherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl or aryl;    -   A₁ and A₂ together are —CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or        —CH(halogen)-CH(OH)—; and    -   X¹ is hydrogen, halogen, NH—OR, NH—O—(CH₂)_(n)COOR, or        NH—O—(CH₂)_(n)CON(R)₂.

In still another subembodiment of formula III, a compound is providedwherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl or aryl;    -   A₁ and A₂ together are 1,2-oxirane or 1,2-cyclopropadiyl; and    -   X¹ together with X² represent a covalent bond.

In another subembodiment of formula III, a compound is provided wherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   A₁ and A₂ together are 1,2-oxirane or 1,2-cyclopropadiyl; and    -   X¹ is hydrogen, halogen, NH—OR, NH—O—(CH₂)_(n)COOR,        NH—O—(CH₂)_(n)CON(R)₂.

In a fourth principal embodiment of the invention, a compound of formulaIV, a tautomer thereof, or a pharmaceutically acceptable salt, solvate,ester or prodrug thereof, is provided:

wherein:

-   -   R, R¹, R², R³, R⁴, R⁵, A₁ and A₂ are as defined above for        formula I; and    -   R⁶ is hydrogen, OR or N(R)₂; and wherein R can be the same or        different.

In one subembodiment of compound IV, R⁶ is hydrogen.

In another subembodiment of compound IV, R⁶ is O-acetyl orO-trifluoroacetyl.

In another subembodiment of compound IV, R¹ is H, halogen orheterocyclyl.

In still another subembodiment of compound IV, R⁵ is hydrogen, alkyl,lower alkyl, aryl, heteroaryl or arylalkyl.

In another subembodiment of compound IV, A₁ and A₂ together are—CH₂—CH₂— or —CH═CH—.

In still another subembodiment of compound IV, A₁ and A₂ together are—CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or —CH(halogen)-CH(OH)—.

In a further subembodiment of compound IV, A₁ and A₂ together are1,2-cyclopropadiyl, or 1,2-oxirane.

In one subembodiment of formula IV, a compound is provided wherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   R⁶ is hydrogen or OR; and    -   A₁ and A₂ together are —CH═CH—.

In another subembodiment of compound IV:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   R⁶ is hydrogen or OR; and    -   A₁ and A₂ together are —CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or        —CH(halogen)-CH(OH)—.

In still another subembodiment of formula IV, a compound is providedwherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is R⁵ is hydrogen, alkyl, aryl or aralkyl;    -   R⁶ is hydrogen or OR; and    -   A₁ and A₂ together are 1,2-oxirane or 1,2-cyclopropadiyl.

In a fifth principal embodiment of the invention, a compound of formulaV, a tautomer thereof, or a pharmaceutically acceptable salt, solvate,ester or prodrug thereof, is provided:

-   -   wherein R, R¹, R², R⁴, R³, R⁵, A₁ and A₂ are as defined above        for formula I; and    -   R⁶ is (CH₂)_(n)C(O)OR, —(CH₂)_(n)C(O)N(R)₂, or        —(CH₂)_(n)C(O)N(R)—OR; where n is 0, 1, 2 or 3; and wherein each        R can be the same or different.

In one subembodiment of compound V, R⁶ is —CH₂C(O)N(Me)OMe.

In one subembodiment of compound V, R¹ is H, halogen or heterocyclyl.

In another subembodiment of compound V, R⁵ is hydrogen, alkyl, loweralkyl, aryl, heteroaryl or arylalkyl.

In another subembodiment of compound V, A₁ and A₂ together are —CH═CH—.

In still another subembodiment of compound IV, A₁ and A₂ together are—CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or CH(halogen)-CH(OH)—.

In a further subembodiment of compound V, A₁ and A₂ together are1,2-cyclopropadiyl, or 1,2-oxirane.

In one subembodiment of formula V, a compound is provided wherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl; and    -   A₁ and A₂ together are —CH═CH— or —CH₂—CH₂—.

In another subembodiment of formula V, a compound is provided wherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl or aryl; and    -   A₁ and A₂ together are —CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or        —CH(halogen)-CH(OH)—.

In still another subembodiment of formula V, a compound is providedwherein:

-   -   R¹ is H, Cl or heterocyclyl;    -   R² and R⁴ are independently OH or OR;    -   R⁵ is hydrogen, alkyl, aryl or aralkyl; and    -   A₁ and A₂ together are 1,2-oxirane or 1,2-cyclopropadiyl.

In specific embodiments of the present invention, the compoundspresented in Table 1, tautomers thereof, or pharmaceutically acceptablesalts, solvates, esters or prodrugs thereof, are provided:

TABLE 1 Compound Designation Structure (R)-2-85e(Ex. 134-I)

(R)-2-85f

(S)-2-85(Ex. 134-F)

(R)-2-85a(Ex. 134-G)

(S)-2-85a(Ex. 134-H)

(S)-145a

(R)-2-103d(Ex. 134-N)

(R)-2-103a(Ex. 134-L)

(S)-2-103a(Ex. 134 M)

(R)-2-103e(Ex. 134-O)

(S)-2-103

(R)-2-103f

2-121g

d-2-121g

(S)-d-2-121(Ex. 134-A)

(R)-d-2-121d(Ex. 134-D)

(R)-d-2-121a(Ex. 134-B)

(R)-2-152a

(S)-2-152a

(S)-2-152a-1X = Cl

(S)-d-2-121a(Ex. 134C)

(S)-2-145aX = Cl

(R)-2-149-1X = Cl

(R)-2-151

(R)-2-151-1X = Cl

(R)-2-151a-1X = Cl

(R)-2-142-1X = Cl(Ex. 136-A)

(R)-2-144-1X = Cl

(R)-2-146-1X = Cl

(R)-2-147-1X = Cl

(R)-trans-2-150-1(Ex. 125)

(R)-2-150c-1X = Cl

(R)-2-150e

Z-(R)-2-155-1

Z-isomer E-(R)-2-155-1

E-isomer (R)-2-147-4X = Cl

(R)-2-153

(S)-2-150-1X = Cl

(S)-cis-2-150

(R)-2-150f

(R)-cis-2-150e

(S)-trans-2-150

(S)-2-150a-1X = Cl

(S)-2-150a

(R)-2-150a

(R)-trans-2-150e

E-(R)-2-155-1

2-147-3

(R)-2-147-3X = Cl

(R)-2-147-2

(R)-cis-2-150d

(R)-trans-2-150d

Z-(R)-2-158-1X = Cl

(R)-2-112d(Ex. 133-D)

(S)-2-112

(R)-2-112a(Ex. 3)

(S)-2-112a(Ex. 133-C)

(R)-2-120d(Ex. 133-I)

(R)-2-120a(Ex. 133-G)

2-185g

2-103g

(R)-2-112(Ex. 105)

(S)-2-120aa(Ex. 133-H)

(R)-2-112e(Ex. 133-J)

(R)-2-112fX = Cl

(R)-2-128

Z-(R)-2-157-1

Z-isomer E-(R)-2-157-1

E-isomer (R)-2-154-1

(R)-2-112

(S)-2-120(Ex. 133-F)

(R)-2-141-2X = Cl

(R)-2-141X = Cl

(R)-2-143X = Cl

2-154-2 bisR¹ = Cl

2-158-2 bisR¹ = Cl

7-1R¹ = HR⁴X = O

2-156-2

2-156-3

(R) 2-147-5

2-154-4

2-154-6

2-154-5

2-154-7

2-154-8

2-154-3

2-154-9

2-170

2-171

2-172-1X = H

2-172-4

2-172-6

2-172-7

2-172-2,

2-172-9

(R)-2-154d-1

(R)-2-154d-5

(R)-2-154d-3

(R)-2-154a-1

(R)-2-154a-5

(R)-2-154a-3

(S)-2-154a-1

(R)-2-154a-5

(S)-2-154a-3

(R)-2-154d-5

(R)-2-154d-3

(R)-2-172a-1

(R)-2-172d-1

(R)-2-172a-5

(R)-2-172a-2

(S)-2-172a-1

(S)-2-173a-5

(S)-2-172a-2

2-155-4

2-155-7

2-144

(R)-2-144a

2-141d

2-174d

2-174

(S)-2-142-1X = Cl

(S)-2-142

2-172-4

2-172-1

(R)-2-120f

2-172-7

2-172-2

(R)-2-120

(S)-2-174

(R)-2-154d-3

(R)-172-2

2-43b

Z-2-a1

Z-isomer E-2a1

E-izomer Z-2-a1-1

Z-isomer E-2-a1

E-isomer 2a-19

(R)-p-2-150

(R)-2-175

2-163

2-164

2-165

2-166

2-167

2-168

2-169

2-a2

2-a3

2-a4

2-a5

2-a6

2-a7

2-a8

2-a9

2-a10

2-a11

2-a12

2-a12

2-a13

E isomer 2-a15

2-a16

2-a17

2-a18

Pharmaceutically Acceptable Salts and Prodrugs

The terms “pharmaceutically acceptable salt” and “prodrug” are usedthroughout the specification to describe any pharmaceutically acceptableform (such as a salt, an ester, a phosphate ester, salt of an ester or arelated group) of a compound which, upon administration to a patient,provides the compound described in the specification. In cases wherecompounds are sufficiently basic or acidic to form stable nontoxic acidor base salts, administration of the compounds as salts may beappropriate. The term pharmaceutically acceptable salts or complexesrefers to salts or complexes that retain the desired biological activityof the compounds of the present invention and exhibit minimal undesiredtoxicological effects.

Nonlimiting examples of such salts are (a) acid addition salts formedwith inorganic acids such as sulfate, nitrate, bicarbonate, andcarbonate salts (for example, hydrochloric acid, hydrobromic acid,sulfuric acid, phosphoric acid, nitric acid, and the like), and saltsformed with organic acids including tosylate, methanesulfonate, acetate,citrate, malonate, tartarate, succinate, benzoate, ascorbate,α-ketoglutarate, and α-glycerophosphate salts, such as acetic acid,oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid,benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid,naphthalenesulfonic acid, naphthalenedisulfonic acid, andpolygalacturonic acid; (b) base addition salts formed with metal cationssuch as zinc, calcium, bismuth, barium, magnesium, aluminum, copper,cobalt, nickel, cadmium, sodium, potassium, lithium and the like, orwith a cation formed from ammonia, N,N-dibenzylethylenediamine,D-glucosamine, tetraethylammonium, or ethylenediamine; or (c)combinations of (a) and (b); e.g., a zinc tannate salt or the like. Alsoincluded in this definition are pharmaceutically acceptable quaternarysalts known by those skilled in the art, which specifically include thequaternary ammonium salt of the formula —NR⁺A⁻, wherein R is as definedabove and A is a counterion, including chloride, bromide, iodide,—O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, orcarboxylate (such as benzoate, succinate, acetate, glycolate, maleate,malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate,benzyloate, and diphenylacetate).

Pharmaceutically acceptable salts may be obtained using standardprocedures well known in the art, for example by reacting a sufficientlybasic compound such as an amine with a suitable acid affording aphysiologically acceptable anion.

Pharmaceutically acceptable “prodrugs” refer to a compound that ismetabolized, for example hydrolyzed or oxidized, in the host to form thecompound of the present invention. Typical examples of prodrugs includecompounds that have biologically labile protecting groups on afunctional moiety of the active compound. Prodrugs include compoundsthat can be oxidized, reduced, aminated, deaminated, hydroxylated,dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated,acylated, deacylated, phosphorylated, dephosphorylated to produce theactive compound. For example, a suitable prodrug may be an ester or anamide of a carboxylic acid that is hydrolyzed to form the acid.Non-limiting examples of prodrugs include but are not limited to alkylor aralkyl esters or amides, including methyl, ethyl, propyl, benzyl andsubstituted benzyl esters or amides. Prodrugs also comprise phosphateesters of the compounds.

Stereoisomerism and Polymorphism

Compounds of the present invention having a chiral center may exist inand be isolated in optically active and racemic forms. The presentinvention encompasses any racemic, optically-active, diastereomeric,polymorphic, or stereoisomeric form, or mixtures thereof, of a compoundof the invention, which possess the useful properties described herein.

In one embodiment, the compounds are prepared in optically active formby asymmetric synthesis using the processes described herein orsynthetic transformations known to those skilled in the art.

Examples of methods to obtain optically active materials are known inthe art, and include at least the following.

i) physical separation of crystals—a technique whereby macroscopiccrystals of the individual enantiomers are manually separated. Thistechnique can be used if crystals of the separate enantiomers exist,i.e., the material is a conglomerate, and the crystals are visuallydistinct;

ii) simultaneous crystallization—a technique whereby the individualenantiomers are separately crystallized from a solution of the racemate,possible only if the latter is a conglomerate in the solid state;

iii) enzymatic resolutions—a technique whereby partial or completeseparation of a racemate by virtue of differing rates of reaction forthe enantiomers with an enzyme;

iv) enzymatic asymmetric synthesis—a synthetic technique whereby atleast one step of the synthesis uses an enzymatic reaction to obtain anenantiomerically pure or enriched synthetic precursor of the desiredenantiomer;

v) chemical asymmetric synthesis—a synthetic technique whereby thedesired enantiomer is synthesized from an achiral precursor underconditions that produce asymmetry (i.e., chirality) in the product,which may be achieved using chiral catalysts or chiral auxiliaries;

vi) diastereomer separations—a technique whereby a racemic compound isreacted with an enantiomerically pure reagent (the chiral auxiliary)that converts the individual enantiomers to diastereomers. The resultingdiastereomers are then separated by chromatography or crystallization byvirtue of their now more distinct structural differences and the chiralauxiliary later removed to obtain the desired enantiomer;

vii) first- and second-order asymmetric transformations—a techniquewhereby diastereomers from the racemate equilibrate to yield apreponderance in solution of the diastereomer from the desiredenantiomer or where preferential crystallization of the diastereomerfrom the desired enantiomer perturbs the equilibrium such thateventually in principle all the material is converted to the crystallinediastereomer from the desired enantiomer. The desired enantiomer is thenreleased from the diastereomer;

viii) kinetic resolutions—this technique refers to the achievement ofpartial or complete resolution of a racemate (or of a further resolutionof a partially resolved compound) by virtue of unequal reaction rates ofthe enantiomers with a chiral, non-racemic reagent or catalyst underkinetic conditions;

ix) enantiospecific synthesis from non-racemic precursors—a synthetictechnique whereby the desired enantiomer is obtained from non-chiralstarting materials and where the stereochemical integrity is not or isonly minimally compromised over the course of the synthesis;

x) chiral liquid chromatography—a technique whereby the enantiomers of aracemate are separated in a liquid mobile phase by virtue of theirdiffering interactions with a stationary phase. The stationary phase canbe made of chiral material or the mobile phase can contain an additionalchiral material to provoke the differing interactions;

xi) chiral gas chromatography—a technique whereby the racemate isvolatilized and enantiomers are separated by virtue of their differinginteractions in the gaseous mobile phase with a column containing afixed non-racemic chiral adsorbent phase;

xii) extraction with chiral solvents—a technique whereby the enantiomersare separated by virtue of preferential dissolution of one enantiomerinto a particular chiral solvent; or

xiii) transport across chiral membranes—a technique whereby a racemateis placed in contact with a thin membrane barrier. The barrier typicallyseparates two miscible fluids, one containing the racemate, and adriving force such as concentration or pressure differential causespreferential transport across the membrane barrier. Separation occurs asa result of the non-racemic chiral nature of the membrane which allowsonly one enantiomer of the racemate to pass through.

DEFINITIONS

Whenever a term in the specification is identified as a range (i.e. C₁₋₄alkyl), the range independently refers to each element of the range. Asa non-limiting example, C₁₋₄ alkyl means, independently, C₁, C₂, C₃ orC₄ alkyl. Similarly, when one or more substituents are referred to asbeing “independently selected from” a group, this means that eachsubstituent can be any element of that group, and any combination ofthese groups can be separated from the group. For example, if R¹ and R²can be independently selected from X, Y and Z, this separately includesthe groups R¹ is X and R² is X; R¹ is X and R² is Y; R¹ is X and R² isZ; R¹ is Y and R² is X; R¹ is Y and R² is Y; R¹ is Y and R² is Z; R¹ isZ and R² is X; R¹ is Z and R² is Y; and R¹ is Z and R² is Z.

The term “alkyl” as used herein, unless otherwise specified, refers to asaturated straight, branched, or cyclic, primary, secondary, or tertiaryhydrocarbon, including but not limited to groups with C₁ to C₁₀.

The term “lower alkyl” refers to a saturated straight, branched, orcyclic, primary, secondary, or tertiary hydrocarbon, including groupswith C₁ to C₄, and if appropriate a cyclic alkyl group (for examplecyclopropyl).

Illustrative examples of alkyl groups are methyl, ethyl, propyl,isopropyl, cyclopropyl, butyl, secbutyl, isobutyl, tertbutyl,cyclobutyl, 1-methylbutyl, 1,1-dimethylpropyl, pentyl, cyclopentyl,isopentyl, neopentyl, cyclopentyl, hexyl, isohexyl, and cyclohexyl.Unless otherwise specified, the alkyl group can be unsubstituted orsubstituted with one or more moieties selected from the group consistingof alkyl, halo, haloalkyl, hydroxyl, carboxyl, acyl, acyloxy, amino,amido, carboxyl derivatives, alkylamino, dialkylamino, arylamino,alkoxy, aryloxy, nitro, cyano, thiol, imine, sulfonic acid, sulfate,sulfonyl, sulfanyl, sulfinyl, sulfamonyl, ester, carboxylic acid, amide,phosphonyl, phosphinyl, phosphoryl, phosphine, thioester, thioether,acid halide, anhydride, oxime, hydrozine, carbamate, phosphonic acid,phosphate, phosphonate, or any other viable functional group that doesnot inhibit the pharmacological activity of this compound, eitherunprotected, or protected as necessary, as known to those skilled in theart, for example, as taught in Greene, et al., Protective Groups inOrganic Synthesis, John Wiley and Sons, Second Edition, 1991.

The term “halo” or “halogen”, as used herein, includes chloro, bromo,iodo, and fluoro.

The term “chiral” as used herein includes a compound that has theproperty that it is not superimposable on its mirror image.

The term “tautomer” as used herein refers to alternate structures whichare recognized in the art to be in equilibrium with the depictedstructure. For example, the enol structure below is a tautomer of theketone structure and recognized to be in equilibrium with the ketonestructure.

As used herein, the term “solvate” or “pharmaceutically acceptablesolvate,” is a solvate formed from the association of one or moresolvent molecules to one or more molecules of a compound of any one offormulas I, II, III, IV or V or the compounds depicted in Table 1. Theterm solvate includes hydrates (e.g., hemi-hydrate, mono-hydrate,dihydrate, trihydrate, tetrahydrate, and the like).

The term “alkylthio” refers to a straight or branched chain alkylsulfideof the number of carbons specified, such as for example, C₁₋₄alkylthio,ethylthio, —S-alkyl, —S-alkenyl, —S-alkynyl, etc.

The terms “alkylamino” or “arylamino” refer to an amino group that hasone or two alkyl or aryl substituents, respectively. Unless otherwisespecifically stated in this application, when alkyl is a suitablemoiety, then it is a lower alkyl, whether substituted or unsubstituted.

The term “alkylsulfonyl” means a straight or branched alkylsulfone ofthe number of carbon atoms specified, as for example, C₁₋₆ alkylsulfonylor methylsulfonyl.

The term “alkoxycarbonyl” refers to a straight or branched chain esterof a carboxylic acid derivative of the number of carbon atoms specified,such as for example, a methoxycarbonyl, MeOCO—.

As used herein, the term “nitro” means —NO₂; the term “sulfhydryl” means—SH; and the term “sulfonyl” means —SO₂.

The terms “alkenyl” and “alkynyl” refer to alkyl moieties, includingboth substituted and unsubstituted forms wherein at least one saturatedC—C bond is replaced by a double or triple bond. Thus, C₂₋₆ alkenyl maybe vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl,3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl,2-hexenyl, 3-hexenyl, 4-hexenyl, or 5-hexenyl. Similarly, C₂₋₆ alkynylmay be ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl,1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl,3-hexynyl, 4-hexynyl, or 5-hexynyl.

The term “alkylene” includes a saturated, straight chain, divalent alkylradical of the formula —(CH₂)_(n)—, wherein “n” may be any whole integerfrom 1 to 10.

“Alkyl”, “alkoxy”, “alkenyl”, “alkynyl”, etc., includes both straightchain and branched groups. However, reference to an individual radicalsuch as “propyl” embraces only that straight-chain radical, whereas abranched chain isomer such as “isopropyl” is specifically termed such.

The term “aryl” as used herein and unless otherwise specified refers toany stable monocyclic, bicyclic, or tricyclic carbon ring of up to 8members in each ring, wherein at least one ring is aromatic as definedby the Huckel 4n+2 rule, and especially phenyl, biphenyl, or naphthyl.The term includes both substituted and unsubstituted moieties. The arylgroup can be substituted with any described moiety, including but notlimited to one or more moieties selected from the group consisting ofhalogen (fluoro, chloro, bromo or iodo), hydroxyl, amino, azido,alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid,sulfate, phosphonic acid, phosphate, or phosphonate, either protected orunprotected as necessary, as known to those skilled in the art, forexample, as taught in Greene et al., Protective Groups in OrganicSynthesis, John Wiley & Sons, 3^(rd) Ed., 1999.

The term “alkaryl” or “alkylaryl” refers to an alkyl group with an arylsubstituent or an alkyl group linked to the molecule through an arylgroup as defined herein. The term “aralkyl” or “arylalkyl” refers to anaryl group substituted with an alkyl substituent or linked to themolecule through an alkyl group as defined above.

The term “cycloalkyl” includes a ring of C₃₋₈, including but not limitedto cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl andcyclooctyl.

The term “alkoxy” means a straight or branched chain alkyl group havingan attached oxygen radical, the alkyl group having the number of carbonsspecified or any number within this range. For example, a “—O-alkyl”,C₁₋₄ alkoxy, methoxy, etc.

The term “acyl” includes a group of the formula C(O)R′, wherein R′ is anstraight, branched, or cyclic alkyl (including lower alkyl), carboxylateresidue of an amino acid, aryl including phenyl, heteroaryl, alkaryl,aralkyl including benzyl, alkoxyalkyl including methoxymethyl,aryloxyalkyl such as phenoxymethyl; or substituted alkyl (includinglower alkyl), aryl including phenyl optionally substituted with chloro,bromo, fluoro, iodo, C₁ to C₄ alkyl or C₁ to C₄ alkoxy, sulfonate esterssuch as alkyl or aralkyl sulphonyl including methanesulfonyl, the mono,di or triphosphate ester, trityl or monomethoxy-trityl, substitutedbenzyl, alkaryl, aralkyl including benzyl, alkoxyalkyl includingmethoxymethyl, aryloxyalkyl such as phenoxymethyl. Aryl groups optimallycomprise a phenyl group. In nonlimiting embodiments, acyl groups includeacetyl, trifluoroacetyl, methylacetyl, cyclopropylacetyl,cyclopropyl-carboxy, propionyl, butyryl, isobutyryl, hexanoyl,heptanoyloctanoyl, neo-heptanoyl, phenylacetyl,2-acetoxy-2-phenylacetyl, diphenylacetyl,α-methoxy-α-trifluoromethyl-phenylacetyl, bromoacetyl,2-nitro-benzeneacetyl, 4-chloro-benzeneacetyl,2-chloro-2,2-diphenylacetyl, 2-chloro-2-phenylacetyl, trimethylacetyl,chlorodifluoroacetyl, perfluoroacetyl, fluoroacetyl,bromodifluoroacetyl, methoxyacetyl, 2-thiopheneacetyl,chlorosulfonylacetyl, 3-methoxyphenylacetyl, phenoxyacetyl,tert-butylacetyl, trichloroacetyl, monochloro-acetyl, dichloroacetyl,7H-dodecafluoro-heptanoyl, perfluoro-heptanoyl,7H-dodeca-fluoroheptanoyl, 7-chlorododecafluoro-heptanoyl,7-chloro-dodecafluoro-heptanoyl, 7H-dodecafluoroheptanoyl,7H-dodeca-fluoroheptanoyl, nona-fluoro-3,6-dioxa-heptanoyl,nonafluoro-3,6-dioxaheptanoyl, perfluoroheptanoyl, methoxybenzoyl,methyl 3-amino-5-phenylthiophene-2-carboxyl,3,6-dichloro-2-methoxy-benzoyl, 4-(1,1,2,2-tetrafluoro-ethoxy)-benzoyl,2-bromo-propionyl, omega-aminocapryl, decanoyl, n-pentadecanoyl,stearyl, 3-cyclopentyl-propionyl, 1-benzene-carboxyl, O-acetylmandelyl,pivaloyl acetyl, 1-adamantane-carboxyl, cyclohexane-carboxyl,2,6-pyridinedicarboxyl, cyclopropane-carboxyl, cyclobutane-carboxyl,perfluorocyclohexyl carboxyl, 4-methylbenzoyl, chloromethyl isoxazolylcarbonyl, perfluorocyclohexyl carboxyl, crotonyl,1-methyl-1H-indazole-3-carbonyl, 2-propenyl, isovaleryl,1-pyrrolidinecarbonyl, 4-phenylbenzoyl.

The term “acylamino” includes a group having a structure of“—N(R′)—C(═O)—R′”, wherein each R′ is independently as defined above.

The term “ester” includes a group of the structure “—C(═O)—O—R′” or“—O—C(═O)—R′”, wherein R′ is an straight, branched, or cyclic alkyl(including lower alkyl), carboxylate residue of an amino acid, arylincluding phenyl, heteroaryl, alkaryl, aralkyl including benzyl,alkoxyalkyl including methoxymethyl, aryloxyalkyl such as phenoxymethyl;or substituted alkyl (including lower alkyl), aryl including phenyloptionally substituted with chloro, bromo, fluoro, iodo, C₁ to C₄ alkylor C₁ to C₄ alkoxy, sulfonate esters such as alkyl or aralkyl sulphonylincluding methanesulfonyl, the mono, di or triphosphate ester, trityl ormonomethoxy-trityl, substituted benzyl, alkaryl, aralkyl includingbenzyl, alkoxyalkyl including methoxymethyl, aryloxyalkyl such asphenoxymethyl. Aryl groups optimally comprise a phenyl group.

The term “heteratom” includes an atom other than carbon or hydrogen inthe structure of a heterocyclic compound, nonlimiting examples of whichare nitrogen, oxygen, sulfur, phosphorus or boron.

The term “carbonyl” or “includes a group of the structure “—C(═O)—X—R′”or “X—C(═O)—R′”, where X is O, S, or a bond, and each R is independentlyas defined above for “ester”.

The term “heterocycle”, “heterocyclyl”, or “heterocyclic” as used hereinincludes non-aromatic ring systems having four to fourteen members,preferably five to ten, in which one or more ring carbons, preferablyone to four, are each replaced by a heteroatom. Examples of heterocyclicrings include 3-1H-benzimidazol-2-one,(1-substituted)-2-oxo-benzimidazol-3-yl, 2-tetrahydro-furanyl,3-tetrahydrofuranyl, 2-tetrahydropyranyl, 3-tetrahydropyranyl,4-tetra-hydropyranyl, [1,3]-dioxalanyl, [1,3]-dithiolanyl,[1,3]-dioxanyl, 2-tetra-hydro-thiophenyl, 3-tetrahydrothiophenyl,2-morpholinyl, 3-morpholinyl, 4-morpholinyl, 2-thiomorpholinyl,3-thiomorpholinyl, 4-thiomorpholinyl, 1-pyrrolidinyl, 2-pyrrolidinyl,3-pyrrolidinyl, 1-piperazinyl, 2-piperazinyl, 1-piperidinyl,2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 4-thiazolidinyl,diazolonyl, N-substituted diazolonyl, 1-phthalimidinyl, benzoxanyl,benzopyrrolidinyl, benzopiperidinyl, benzoxolanyl, benzothiolanyl, andbenzothianyl. Also included within the scope of the term “heterocyclyl”or “heterocyclic”, as it is used herein, is a group in which anon-aromatic heteroatom-containing ring is fused to one or more aromaticor non-aromatic rings, such as in an indolinyl, chromanyl,phenanthridinyl, or tetrahydroquinolinyl, where the radical or point ofattachment is on the non-aromatic heteroatom-containing ring. The term“heterocycle”, “heterocyclyl”, or “heterocyclic” whether saturated orpartially unsaturated, also refers to rings that are optionallysubstituted.

The term “heteroaryl”, used alone or as part of a larger moiety as in“heteroaralkyl” or “heteroarylalkoxy”, refers to heteroaromatic ringgroups having five to fourteen members. Examples of heteroaryl ringsinclude 2-furanyl, 3-furanyl, 3-furazanyl, N-imidazolyl, 2-imidazolyl,4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl,2-oxadiazolyl, 5-oxadiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl,1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 1-pyrazolyl, 2-pyrazolyl,3-pyrazolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl,5-pyrimidyl, 3-pyridazinyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,5-tetrazolyl, 2-triazolyl, 5-triazolyl, 2-thienyl, 3-thienyl,carbazolyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl,quinolinyl, benzotriazolyl, benzothiazolyl, benzooxazolyl,benzimidazolyl, isoquinolinyl, indazolyl, isoindolyl, acridinyl, andbenzoisoxazolyl. Also included within the scope of the term“heteroaryl”, as it is used herein, is a group in which a heteroatomicring is fused to one or more aromatic or nonaromatic rings where theradical or point of attachment is on the heteroaromatic ring. Examplesinclude tetrahydroquinolinyl, tetrahydroisoquinolinyl, andpyrido[3,4-d]pyrimidinyl. The term “heteroaryl” also refers to ringsthat are optionally substituted. The term “heteroaryl” may be usedinterchangeably with the term “heteroaryl ring” or the term“heteroaromatic”.

The term “amino” as used herein unless otherwise specified, includes amoiety represented by the structure “—N(R)₂”, and includes primary,secondary and tertiary amines optionally substituted by alkyl, aryl,heterocyclyl, and/or sulfonyl groups. Thus (R)₂ may represent twohydrogen atoms, two alkyl moieties, or one hydrogen and one alkylmoiety.

The term “amido” as used herein includes an amino-substituted carbonyl,while the term “amidino” means a group having the structure“—C(═NH)—NH₂”.

The term “quaternary amine” as used herein includes quaternary ammoniumsalts that have a positively charged nitrogen. They are formed by thereaction between a basic nitrogen in the compound of interest and anappropriate quaternizing agent such as, for example, methyliodide orbenzyliodide. Appropriate counterions accompanying a quaternary amineinclude acetate, trifluoroacetate, chloro, bromo and iodo ions.

The term “substituted” includes multiple degrees of substitution by oneor more named substituents such as, for example, halo, hydroxyl, thio,alkyl, alkenyl, alkynyl, nitro, cyano, azido, amino, carboxamido, etc.Where multiple substituent possibilities exist, the compound can besubstituted by one or more of the disclosed or claimed substituentgroups, independently from one another, and taken singly or plurally.

The term “protected” as used herein and unless otherwise defined refersto a group that is added to an oxygen, nitrogen, or phosphorus atom toprevent its further reaction or for other purposes. A wide variety ofoxygen and nitrogen protecting groups are known to those skilled in theart of organic synthesis.

The term “protecting group” as used herein refers to a group that may beattached to a reactive group, including heteroatoms such as oxygen ornitrogen, to prevent the reactive group from participating in areaction. Any protecting groups taught in Greene, et al., ProtectiveGroups in Organic Synthesis, John Wiley and Sons, Second Edition, 1991may be used. Examples of suitable protecting groups include but are notlimited to alkoxyalkyl groups such as ethoxymethyl and methoxymethyl;silyl protecting groups, such tert-butyldimethyl silyl (TBS),phenyldimethylsilyl, trimethylsilyl (TMS), 2-trimethylsilylethoxymethyl(SEM) and 2-trimethylsilylethyl; and benzyl and substituted benzyl.

It should be understood that the various possible stereoisomers of thegroups mentioned above and herein are within the meaning of theindividual terms and examples, unless otherwise specified. As anillustrative example, “1-methyl-butyl” exists in both (R) and the (S)form, thus, both (R)-1-methyl-butyl and (S)-1-methyl-butyl is covered bythe term “1-methyl-butyl”, unless otherwise specified.

The term “patient” includes human and veterinary subjects.

An “effective amount” is the quantity of compound in which a beneficialoutcome is achieved when the compound is administered to a patient oralternatively, the quantity of compound that possesses a desiredactivity in vivo or in vitro. In the case of proliferative disorders, abeneficial clinical outcome includes reduction in the extent or severityof the symptoms associated with the disease or disorder and/or anincrease in the longevity and/or quality of life of the patient comparedwith the absence of the treatment. For example, for a subject withcancer, a “beneficial clinical outcome” includes a reduction in tumormass, a reduction in the rate of tumor growth, a reduction inmetastasis, a reduction in the severity of the symptoms associated withthe cancer and/or an increase in the longevity of the subject comparedwith the absence of the treatment. The precise amount of compoundadministered to a subject will depend on the type and severity of thedisease or condition and on the characteristics of the patient, such asgeneral health, age, sex, body weight and tolerance to drugs. It willalso depend on the degree, severity and type of proliferative disorder.The skilled artisan will be able to determine appropriate dosagesdepending on these and other factors.

The term “kinase-inhibiting amount” as used herein, refers to an amountof the compound that inhibits a kinase enzyme compared to a control astested by the methods described herein.

The term “HSP 90-inhibiting amount” as used herein, refers to an amountof the compound that inhibits HSP90 compared to a control as tested bythe methods described herein.

The term “biological sample”, as used herein, includes, withoutlimitation, cell cultures or extracts thereof; preparations of an enzymesuitable for in vitro assay; biopsied material obtained from a mammal orextracts thereof; and blood, saliva, urine, feces, semen, tears, orother body fluids or extracts thereof.

The term “cancer” includes, but is not limited to, solid tumors andblood borne tumors and include, but is not limited to, the followingcancers: breast, ovary, cervix, prostate, testis, genitourinary tract,esophagus, larynx, glioblastoma, stomach, skin, keratoacanthoma, lung,epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lungadenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma, thyroid,follicular carcinoma, undifferentiated carcinoma, papillary carcinoma,seminoma, melanoma, sarcoma, bladder carcinoma, liver carcinoma andbiliary passages, kidney carcinoma, myeloid disorders, lymphoiddisorders, Hodgkin's, hairy cells, buccal cavity and pharynx (oral),lip, tongue, mouth, pharynx, small intestine, colon-rectum, largeintestine, rectum, brain and central nervous system, and leukemia. Theterm “cancer” includes primary cancer, cancers secondary to treatment,and metastatic cancers.

The term “pharmaceutically acceptable carrier” refers to a non-toxiccarrier, adjuvant, or vehicle that may be administered to a patient,together with a compound of this invention, and which does not destroythe pharmacological activity thereof.

The terms “GSK-3-mediated disease, or “GSK-3-mediated condition”, asused herein, mean any disease or other deleterious condition or state inwhich GSK-3 is known to play a role. Such diseases or conditionsinclude, without limitation, diabetes, Alzheimer's disease, Huntington'sDisease, Parkinson's Disease, AIDS-associated dementia, amyotrophiclateral sclerosis (AML), multiple sclerosis (MS), schizophrenia,cardiomycete hypertrophy, reperfusion/ischemia, and baldness.

The terms “CDK-2-mediated disease” or CDK-2-mediated condition”, as usedherein, mean any disease or other deleterious condition in which CDK-2is known to play a role. The terms “CDK-2-mediated disease” or“CDK-2-mediated condition” also mean those diseases or conditions thatare alleviated by treatment with a CDK-2 inhibitor. Such conditionsinclude, without limitation, cancer, Alzheimer's disease, restenosis,angiogenesis, glomerulonephritis, cytomegalovirus, HIV, herpes,psoriasis, atherosclerosis, alopecia, and autoimmune diseases such asrheumatoid arthritis, such as are described for example in Fischer, P.M. and Lane, D. P., Current Medicinal Chemistry, 7, 1213-1245 (2000);Mani, S., Wang, C., Wu, K., Francis, R. and Pestell, R., Exp. Opin.Invest. Drugs, 9, 1849 (2000); Fry, D. W. and Garrett, M. D., CurrentOpinion in Oncologic, Endocrine & Metabolic Investigational Drugs, 2,40-59 (2000).

The terms “ERK-mediated disease” or “ERK-mediated condition”, as usedherein mean any disease or other deleterious condition in which ERK mayplay a role. The terms “ERK-2-mediated disease” or “ERK-2-mediatedcondition” also mean those diseases or conditions that are alleviated bytreatment with a ERK-2 inhibitor. Such conditions include, withoutlimitation, cancer, stroke, diabetes, hepatomegaly, cardiovasculardisease including cardiomegaly, Alzheimer's disease, cystic fibrosis,viral disease, autoimmune diseases, atherosclerosis, restenosis,psoriasis, allergic disorders including asthma, inflammation,neurological disorders and hormone-related diseases. ERK-2 proteinkinase and its implication in various diseases has been described forexample in Bokemeyer et al. 1996, Kidney Int. 49, 1187; Anderson et al.,1990, Nature 343, 651; Crews et al., 1992, Science 258, 478; Bjorbaek etal., 1995, J. Biol. Chem. 270, 18848; Rouse et al., 1994, Cell 78, 1027;Raingeaud et al., 1996, Mol. Cell. Biol. 16, 1247; Raingeaud et al.1996; Chen et al., 1993 Proc. Natl. Acad. Sci. USA 90, 10952; Oliver etal., 1995, Proc. Soc. Exp. Biol. Med. 210, 162; Moodie et al., 1993,Science 260, 1658; Frey and Mulder, 1997, Cancer Res. 57, 628; Sivaramanet al., 1997, J. Clin. Invest. 99, 1478; Whelchel et al., 1997, Am. J.Respir. Cell Mol. Biol. 16, 589.

The terms “AKT-mediated disease” or “AKT-mediated condition”, as usedherein, mean any disease or other deleterious condition in which AKT isknown to play a role. The terms “AKT-mediated disease” or “AKT-mediatedcondition” also mean those diseases or conditions that are alleviated bytreatment with a AKT inhibitor. AKT-mediated diseases or conditionsinclude, but are not limited to, proliferative disorders, cancer, andneurodegenerative disorders. The association of AKT, also known asprotein kinase B, with various diseases has been described for examplein Khwaja, A., Nature, pp. 33-34, 1990; Zang, Q. Y., et al, Oncogene, 192000; Kazuhiko, N., et al, The Journal of Neuroscience, 20 2000.

The terms “Src-mediated disease” or “Src-mediated condition”, as usedherein mean any disease or other deleterious condition in which Src isknown to play a role. The terms “Src-mediated disease” or “Src-mediatedcondition” also mean those diseases or conditions that are alleviated bytreatment with a Src inhibitor. Such conditions include, withoutlimitation, hypercalcemia, osteoporosis, osteoarthritis, cancer,symptomatic treatment of bone metastasis, and Paget's disease. Srcprotein kinase and its implication in various diseases has beendescribed for example in Soriano, Cell, 69, 551 (1992); Soriano et al.,Cell, 64, 693 (1991); Takayanagi, J. Clin. Invest., 104, 137 (1999);Boschelli, Drugs of the Future 2000, 25(7), 717, (2000); Talamonti, J.Clin. Invest., 91, 53 (1993); Lutz, Biochem. Biophys. Res. 243, 503(1998); Rosen, J. Biol. Chem., 261, 13754 (1986); Bolen, Proc. Natl.Acad. Sci. USA, 84, 2251 (1987); Masaki, Hepatology, 27, 1257 (1998);Biscardi, Adv. Cancer Res., 76, 61 (1999); Lynch, Leukemia, 7, 1416(1993); Wiener, Clin. Cancer Res., 5, 2164 (1999); Staley, Cell GrowthDiff., 8, 269 (1997).

The terms “Lck-mediated disease” or “Lck-mediated condition”, as usedherein, mean any disease state or other deleterious condition in whichLck is known to play a role. The terms “Lck-mediated disease” or“Lck-mediated condition” also mean those diseases or conditions that arealleviated by treatment with an Lck inhibitor. Lck-mediated diseases orconditions include, but are not limited to, autoimmune diseases such astransplant rejection, allergies, rheumatoid arthritis, and leukemia. Theassociation of Lck with various diseases has been described for examplein Molina et al., Nature, 357, 161 (1992).

The terms “Abl-mediated disease” or “Abl-mediated condition”, as usedherein, mean any disease state or other deleterious condition in whichAbl is known to play a role. The terms “Abl-mediated disease” or“Abl-mediated condition” also mean those diseases or conditions that arealleviated by treatment with an Abl inhibitor. Abl-mediated diseases orconditions include, but are not limited to, leukemias, particularlychronic myeloid leukemia. The association of Abl with various diseaseshas been described for example in Druker, et al., N. Engl. J. Med. 2001,344, 1038-1042.

The terms “cKit-mediated disease” or “cKit-mediated condition”, as usedherein, mean any disease state or other deleterious condition in whichcKit is known to play a role. The terms “cKit-mediated disease” or“cKit-mediated condition” also mean those diseases or conditions thatare alleviated by treatment with an cKit inhibitor. cKit-mediateddiseases or conditions include, but are not limited to,mastocytosis/mast cell leukemia, gastrointestinal stromal tumor,sinonasal natural killer/T-cell lymphoma, seminoma/dysgerminoma, throidcarcinoma, samll-cell lung carcinoma, malignant melanoma, adenoid cysticcarcinoma, ovarian carcinoma, acute myelogenious leukemia, anaplasticlarge-cell lymphoma, angiosarcoma, endometrial carcinom, pediatricT-cell ALL/lymphoma, breast carcinoma and prostate carcinoma. Theassociation of cKit with various diseases has been described for examplein Heinrich, et al., J. Clinical Oncology 2002, 20, 1692-1703.

The terms “Flt3-mediated disease” or “Flt3-mediated condition”, as usedherein, mean any disease state or other deleterious condition in whichFlt3 is known to play a role. The terms “Flt3-mediated disease” or“Flt3-mediated condition” also mean those diseases or conditions thatare alleviated by treatment with an Flt3 inhibitor. Flt3-mediateddiseases or conditions include, but are not limited to, acutemyelogenous leukemia, mixed lineage leukemia and acute lymphocyticleukemia. The association of Flt3 with various diseases has beendescribed for example in Sternberg and Licht, Curr. Opin Hematol. 2004,12, 7-13.

The terms “KDR-mediated disease” or “KDR-mediated condition”, as usedherein, mean any disease state or other deleterious condition in whichKDR is known to play a role. The terms “KDR-mediated disease” or“KDR-mediated condition” also mean those diseases or conditions that arealleviated by treatment with an KDR inhibitor. KDR-mediated diseases orconditions include, but are not limited to, carcinoma of the lung,breast, gastrointestinal tract, kidney, bladder, ovary and endometrium,intracranial tumors including glioblatoma multiforme, sporadic capillaryhemangioblastoma, hematological malignancies, including T cell lymphoma,acute lymphoblastic leukemia, Burkitt's lymphoma and promyelocyticleukemia, age-related macular degeneration, herpetic ocular disease,rheumatoid arthritis, cerebral ischemia and endometriosis. Theassociation of KDR with various diseases has been described for examplein Ferrara, Endocrine Reviews 2004, 25, 581-611.

The term “HSP90-mediated disease” or “HSP90-mediated condition” refersto a condition in which HSP90 is known to pay a role. The conditionsinclude but are not limited to inflammatory disorders, abnormal cellularproliferation, autoimmune disorders, ischemia, fibrogenetic disordersincluding but not limited to scleroderma, polymyositis, systemic lupus,rheumatoid arthritis, liver cirrhosis, keloid formation, interstitialnephritis, and pulmonary fibrosis. (Strehlow, WO 02/02123;PCT/US01/20578).

Method of Treatment

The compounds described herein, are particularly useful for thetreatment or prevention of a disorder mediated by kinases or mediated byHSP90. In one embodiment, the compounds described herein, are useful forthe treatment or prevention of a proliferative disorder, includingcancer metastasis. In another embodiment, the compounds describedherein, are useful for the treatment or prevention of an inflammatorydisorder associated by kinases or HSP90.

An aspect of the invention relates to compounds and compositions thatare useful for treating cancer.

Another aspect of the invention relates to the treatment of thefollowing cancers: breast, ovary, cervix, prostate, testis,genitourinary tract, esophagus, larynx, glioblastoma, stomach, skin,keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, smallcell carcinoma, lung adenocarcinoma, bone, colon, adenoma, pancreas,adenocarcinoma, thyroid, follicular carcinoma, undifferentiatedcarcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladdercarcinoma, liver carcinoma and biliary passages, kidney carcinoma,myeloid disorders, lymphoid disorders, Hodgkin's, hairy cells, buccalcavity and pharynx (oral), lip, tongue, mouth, pharynx, small intestine,colon-rectum, large intestine, rectum, brain and central nervous system,and leukemia.

Another aspect of the invention is a method for treating cancercomprising administering an effective amount of a compound of formula I,II, III, IV or V described herein to a patient with cancer.

Angiogenesis is characterized by the proliferation of endothelial cellsto form new blood vessels (often called neovascularization). Inhibitionof mitosis of endothelial cells results in inhibition of angiogenesis.Another aspect of this invention therefore relates to inhibition ofundesirable mitosis, including undesirable angiogenesis. A mammaliandisease characterized by undesirable cell mitosis, as defined herein,includes, but is not limited to, excessive or abnormal stimulation ofendothelial cells (e.g., atherosclerosis), solid tumors and tumormetastasis, benign tumors, for example, hemangiomas, trachomas, andpyogenic granulomas, vascular malfunctions, abnormal wound healing,inflammatory and immune disorders, Bechet's disease, gout or goutyarthritis, abnormal angiogenesis accompanying rheumatoid arthritis, skindiseases, such as psoriasis, diabetic retinopathy and other ocularangiogenic diseases such as retinopathy of prematurity (retrolentalfibroplasic), macular degeneration, corneal graft rejection, neovascularglaucoma and Osler Weber syndrome (Osler-Weber-Rendu disease).

Other undesired angiogenesis involves normal processes includingovulation and implantation of a blastula. The compositions describedabove can be used as a birth control agent by reducing or preventinguterine vascularization required for embryo implantation. Accordingly,the compositions described above can be used to block ovulation andimplantation of a blastula or to block menstruation (induce amenorrhea).

Diseases associated with undesirable mitosis includingneovascularization can be treated according to the present invention.Such diseases include, but are not limited to, ocular neovasculardisease, diabetic retinopathy, retinopathy of prematurity, corneal graftrejection, neovascular glaucoma and retrolental fibroplasias, epidemickeratoconjunctivitis, Vitamin A deficiency, contact lens overwear,atopic keratitis, superior limbic keratitis, pterygium keratitis sicca,Sjögren's syndrome, acne rosacea, phylectenulosis, syphilis,Mycobacteria infections, lipid degeneration, chemical burns, bacterialulcers, fungal ulcers, Herpes simplex infections, Herpes zosterinfections, protozoan infections, Kaposi's sarcoma, Mooren's ulcer,Terrien's marginal degeneration, marginal keratolysis, trauma,rheumatoid arthritis, systemic lupus, polyarteritis, Wegener'ssarcoidosis, Scleritis, Steven-Johnson disease, pemphigoid, radialkeratotomy, and corneal graph rejection.

Other diseases associated with undesirable mitosis includingneovascularization can be treated according to the present invention.Such diseases include, but are not limited to, sickle cell anemia,sarcoid, pseudoxanthoma elasticum, Paget's disease, vein occlusion,artery occlusion, carotid obstructive disease, chronic uveitis/vitritis,Lyme's disease, systemic lupus erythematosis, Eales' disease, Bechet'sdisease, infections causing a retinitis or choroiditis, presumed ocularhistoplasmosis, Best's disease, myopia, optic pits, Stargart's disease,pars planitis, chronic retinal detachment, hyperviscosity syndromes,toxoplasmosis, and post-laser complications. Other diseases include, butare not limited to, diseases associated with rubeosis(neovascularization of the iris and the angle) and diseases caused bythe abnormal proliferation of fibrovascular or fibrous tissue includingall forms of proliferative vitreoretinopathy, whether or not associatedwith diabetes.

Another aspect of the invention relates to the treatment of inflammatorydiseases including, but no limited to, excessive or abnormal stimulationof endothelial cells (e.g., atherosclerosis), solid tumors and tumormetastasis, benign tumors, for example, hemangiomas, acoustic neuromas,trachomas, and pyogenic granulomas, vascular malfunctions, abnormalwound healing, inflammatory and immune disorders, Bechet's disease, goutor gouty arthritis, abnormal angiogenesis accompanying rheumatoidarthritis, skin diseases, such as psoriasis, diabetic retinopathy andother ocular angiogenic diseases such as retinopathy of prematurity(retrolental fibroplasic), macular degeneration, corneal graftrejection, neovascular glaucoma and Osler Weber syndrome(Osler-Weber-Rendu disease). Other undesired angiogenesis involvesnormal processes including ovulation and implantation of a blastula.Accordingly, the compositions described above can be used to blockovulation and implantation of a blastula or to block menstruation(induce amenorrhea).

Another aspect of this invention relates to a method of inhibiting HSP90activity in a patient, comprising administering to a patient aneffective amount of a compound of formula I, II, III, IV or V or apharmaceutically acceptable salt or prodrug thereof. The invention alsoprovides a method for treating a disease that is mediated by HSP90.

Another aspect of this invention relates to a method of inhibitingAurora A activity in a patient, comprising administering to a patient aneffective amount of a compound of formula I, II, III, IV or V or apharmaceutically acceptable salt or prodrug thereof.

Another aspect of this invention relates to a method of treating orpreventing a GSK-3-mediated disease with a GSK-3 inhibitor, comprisingadministering to a patient an effective amount of a compound of formulaI, II, III, IV or V or a pharmaceutically acceptable salt or prodrugthereof.

One aspect of this invention relates to a method of enhancing glycogensynthesis and/or lowering blood levels of glucose in a patient in needthereof, which method comprises administering to the patient atherapeutically effective amount of a compound of formula I, U, III, IVor V or a pharmaceutical composition thereof. This method is especiallyuseful for diabetic patients. Another method relates to inhibiting theproduction of hyperphosphorylated Tau protein, which is useful inhalting or slowing the progression of Alzheimer's disease. Anothermethod relates to inhibiting the phosphorylation of .beta.-catenin,which is useful for treating schizophrenia.

Another aspect of the invention relates to inhibiting GSK-3 activity ina biological sample, which method comprises contacting the biologicalsample with a GSK-3 inhibitor of formula I, II, III, IV or V.

Another aspect of this invention relates to a method of inhibiting GSK-3activity in a patient comprising administering to the patient a compoundof formula I, II, III, IV or V or a composition comprising saidcompound.

Another aspect of this invention relates to a method of treating orpreventing a CDK-2-mediated disease comprising administering to apatient in need of such a treatment a therapeutically effective amountof a compound of formula I, II, III, IV or V or a pharmaceuticalcomposition thereof.

Another aspect of the invention relates to inhibiting CDK-2 activity ina biological sample or a patient, which method comprises administeringto the patient a compound of formula I, II, III, IV or V, or acomposition comprising said compound.

Another aspect of this invention relates to a method of treating orpreventing an ERK-2-mediated diseases comprising administering to apatient in need of such a treatment a therapeutically effective amountof a compound of formula I, II, III, IV or V or a pharmaceuticalcomposition thereof.

Another aspect of the invention relates to inhibiting ERK-2 activity ina biological sample or a patient, which method comprises administeringto the patient a compound of formula I, II, III, IV or V, or acomposition comprising said compound.

Another aspect of this invention relates to a method of treating orpreventing an AKT-mediated diseases comprising administering to apatient in need of such a treatment a therapeutically effective amountof a compound of formula I, II, III, IV or V or a pharmaceuticalcomposition thereof.

Another aspect of the invention relates to inhibiting AKT activity in abiological sample or a patient, which method comprises administering tothe patient a compound of formula I, II, III, IV or V, or a compositioncomprising said compound.

Another aspect of this invention relates to a method of treating orpreventing a Src-mediated disease comprising administering to a patientin need of such a treatment a therapeutically effective amount of acompound of formula I, II, III, IV or V or a pharmaceutical compositionthereof.

Another aspect of the invention relates to inhibiting Src activity in abiological sample or a patient, which method comprises administering tothe patient a compound of formula I, II, III, IV or V, or a compositioncomprising said compound.

Another aspect of this invention relates to a method of treating orpreventing an Lck-mediated disease with an Lck inhibitor, which methodcomprises administering to a patient in need of such a treatment atherapeutically effective amount of a compound of formula I, II, III, IVor V, or a pharmaceutical composition thereof.

Another aspect of the invention relates to inhibiting Lck activity in abiological sample or a patient, which method comprises administering tothe patient a compound of formula I, II, III, IV or V, or a compositioncomprising said compound.

Another aspect of this invention relates to a method of treating orpreventing an Abl-mediated disease with an Abl inhibitor, which methodcomprises administering to a patient in need of such a treatment atherapeutically effective amount of a compound of formula I, II, III, IVor V, or a pharmaceutical composition thereof.

Another aspect of the invention relates to inhibiting Abl activity in abiological sample or a patient, which method comprises administering tothe patient a compound of formula I, II, III, IV or V, or a compositioncomprising said compound.

Another aspect of this invention relates to a method of treating orpreventing a cKit-mediated disease comprising administering to a patientin need of such a treatment a therapeutically effective amount of acompound of formula I, II, I, IV or V, or a pharmaceutical compositionthereof.

Another aspect of the invention relates to inhibiting cKit activity in abiological sample or a patient, which method comprises administering tothe patient a compound of formula I, II, III, IV or V, or a compositioncomprising said compound.

Another aspect of this invention relates to a method of treating orpreventing a Flt3-mediated disease comprising administering to a patientin need of such a treatment a therapeutically effective amount of acompound of formula I, II, III, IV or V, or a pharmaceutical compositionthereof.

Another aspect of the invention relates to inhibiting Flt3 activity in abiological sample or a patient, which method comprises administering tothe patient a compound of formula I, II, I, IV or V, or a compositioncomprising said compound.

Another aspect of this invention relates to a method of treating orpreventing a KDR-mediated disease comprising administering to a patientin need of such a treatment a therapeutically effective amount of acompound of formula I, II, III, IV or V, or a pharmaceutical compositionthereof.

Another aspect of the invention relates to inhibiting KDR activity in abiological sample or a patient, which method comprises administering tothe patient a compound of formula I, II, III, IV or V, or a compositioncomprising said compound.

An amount effective to inhibit protein kinase, is an amount that causesmeasurable inhibition of the kinase activity when compared to theactivity of the enzyme in the absence of an inhibitor. Any method may beused to determine inhibition, such as, for example, the BiologicalTesting Examples described below.

Pharmaceutical Compositions

Mammals, and specifically humans, suffering from a respiratory disordercan be treated by the inhalation, systemic, oral, topical, ortransdermal administration of a composition comprising an effectiveamount of the compounds described herein or a pharmaceuticallyacceptable salt, ester or prodrug thereof, optionally in apharmaceutically acceptable carrier or diluent.

The compounds or compositions are typically administered by oral orinhalation administration. Alternatively, compounds can be administeredsubcutaneously, intravenously, intraperitoneally, intramuscularly,parenterally, orally, submucosally, by inhalation, transdermally via aslow release patch, or topically, in an effective dosage range to treatthe target condition.

An effective dose can be readily determined by the use of conventionaltechniques and by observing results obtained under analogouscircumstances. In determining the effective dose, a number of factorsare considered including, but not limited to: the species of patient;its size, age, and general health; the specific disease involved; thedegree of involvement or the severity of the disease; the response ofthe individual patient; the particular compound administered; the modeof administration; the bioavailability characteristics of thepreparation administered; the dose regimen selected; and the use ofconcomitant medication.

In a separate embodiment, the compounds of the invention are in the formof an inhaled dosage. In this embodiment, the compounds may be in theform of an aerosol suspension, a dry powder or liquid particle form. Thecompounds may be prepared for delivery as a nasal spray or in aninhaler, such as a metered dose inhaler. Pressurized metered-doseinhalers (“MDI”) generally deliver aerosolized particles suspended inchlorofluorocarbon propellants such as CFC-11, CFC-12, or thenon-chlorofluorocarbons or alternate propellants such as thefluorocarbons, HFC-134A or HFC-227 with or without surfactants andsuitable bridging agents. Dry-powder inhalers can also be used, eitherbreath activated or delivered by air or gas pressure such as thedry-powder inhaler disclosed in the Schering Corporation InternationalPatent Application No. PCT/US92/05225, published 7 Jan. 1993 as well asthe Turbuhaler™ (available from Astra Pharmaceutical Products, Inc.) orthe Rotahaler™ (available from Allen & Hanburys) which may be used todeliver the aerosolized particles as a finely milled powder in largeaggregates either alone or in combination with some pharmaceuticallyacceptable carrier e.g. lactose; and nebulizers.

The compounds of the invention may be also administered in specific,measured amounts in the form of an aqueous suspension by use of a pumpspray bottle. The aqueous suspension compositions of the presentinvention may be prepared by admixing the compounds with water and otherpharmaceutically acceptable excipients. The aqueous suspensioncompositions according to the present invention may contain, inter alia,water, auxiliaries and/or one or more of the excipients, such as:suspending agents, e.g., microcrystalline cellulose, sodiumcarboxymethylcellulose, hydroxpropyl-methyl cellulose; humectants, e.g.glycerin and propylene glycol; acids, bases or buffer substances foradjusting the pH, e.g., citric acid, sodium citrate, phosphoric acid,sodium phosphate as well as mixtures of citrate and phosphate buffers;surfactants, e.g. Polysorbate 80; and antimicrobial preservatives, e.g.,benzalkonium chloride, phenylethyl alcohol and potassium sorbate.

Typical systemic dosages for all of the herein described conditions arethose ranging from 0.01 mg/kg to 1500 mg/kg of body weight per day as asingle daily dose or divided daily doses. Preferred dosages for thedescribed conditions range from 0.5-1500 mg per day. A more particularlypreferred dosage for the desired conditions ranges from 5-750 mg perday. Typical dosages can also range from 0.01 to 1500, 0.02 to 1000, 0.2to 500, 0.02 to 200, 0.05 to 100, 0.05 to 50, 0.075 to 50, 0.1 to 50,0.5 to 50, 1 to 50, 2 to 50, 5 to 50, 10 to 50, 25 to 50, 25 to 75, 25to 100, 100 to 150, or 150 or more mg/kg/day, as a single daily dose ordivided daily doses. In one embodiment, the compounds are given in dosesof between about 1 to about 5, about 5 to about 10, about 10 to about 25or about 25 to about 50 mg/kg. Typical dosages for topical applicationare those ranging from 0.001 to 100% by weight of the active compound.

The compounds are conveniently administered in units of any suitabledosage form, including but not limited to one containing from about 7 to3000 mg, from about 70 to 1400 mg, or from about 25 to 1000 mg of activeingredient per unit dosage form. For example, an oral dosage of fromabout 50 to 1000 mg is usually convenient, including in one or multipledosage forms of 50, 100, 200, 250, 300, 400, 500, 600, 700, 800, 900 or1000 mgs. Lower dosages may be preferable, for example, from about10-100 or 1-50 mgs. Also contemplated are doses of 0.1-50 mg, 0.1-20mgs., or 0.1-10 mgs. Furthermore, lower doses may be utilized in thecase of administration by a non-oral route, as for example, by injectionor inhalation.

The compound is administered for a sufficient time period to alleviatethe undesired symptoms and the clinical signs associated with thecondition being treated.

The active compound is included in the pharmaceutically acceptablecarrier or diluent in an amount sufficient to deliver to a patient atherapeutic amount of compound in vivo in the absence of serious toxiceffects. Pharmaceutically acceptable carriers that may be used in thesepharmaceutical compositions are generally known in the art. Theyinclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,solvents, salts or electrolytes, such as protamine sulfate, disodiumhydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zincsalts, silicates, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes, oils,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat. Pharmaceutically accepted vehicles can contain mixtures ofmore than one excipient in which the components and the ratios can beselected to optimize desired characteristics of the formulationincluding but not limited to shelf-life, stability, drug load, site ofdelivery, dissolution rate, self-emulsification, control of release rateand site of release, and metabolism.

Formulations can be prepared by a variety of techniques known in theart. Examples of formulation techniques can be found in literaturepublications and in texts such as “Water-insoluble drug formulation”,edited by Rong Liu, 2000, Interpharm Press.

If administered intravenously, carriers can be physiological saline,bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) orphosphate buffered saline (PBS). Sterile injectable forms of thecompositions of this invention may be aqueous or oleaginous suspension.These suspensions may be formulated according to techniques known in theart using suitable dispersing or wetting agents and suspending agents.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose, any bland fixed oil may be employedincluding synthetic mono- or di-glycerides. Fatty acids, such as oleicacid and its glyceride derivatives are useful in the preparation ofinjectables, as are natural pharmaceutically-acceptable oils, such asolive oil or castor oil, especially in their polyoxyethylated versions.These oil solutions or suspensions may also contain a long-chain alcoholdiluent or dispersant, such as carboxymethyl cellulose or similardispersing agents which are commonly used in the formulation ofpharmaceutically acceptable dosage forms including emulsions andsuspensions. Other commonly used surfactants, such as Tweens, Spans andother surface-active emulsifying agents or bioavailability enhancerswhich are commonly used in the manufacture of pharmaceuticallyacceptable solid, liquid, or other dosage forms may also be used for thepurposes of formulation.

The concentration of active compound in the drug composition will dependon absorption, inactivation, and excretion rates of the drug as well asother factors known to those of skill in the art. It is to be noted thatdosage values will also vary with the severity of the condition to bealleviated. It is to be further understood that for any particularsubject, specific dosage regimens should be adjusted over time accordingto the individual need and the professional judgment of the personadministering or supervising the administration of the compositions, andthat the dosage ranges set forth herein are exemplary only and are notintended to limit the scope or practice of the claimed composition. Theactive ingredient may be administered at once, or may be divided into anumber of smaller doses to be administered at varying intervals of time.

One mode of administration of the active compound for systemic deliveryis oral. Oral compositions will generally include an inert diluent or anedible carrier. They may be enclosed in gelatin capsules or compressedinto tablets. For the purpose of oral therapeutic administration, theactive compound can be incorporated with excipients and used in the formof tablets, troches, or capsules. Pharmaceutically compatible bindingagents, and/or adjuvant materials can be included as part of thecomposition.

The tablets, pills, capsules, troches and the like can contain any ofthe following ingredients, or compounds of a similar nature: a bindersuch as microcrystalline cellulose, gum tragacanth or gelatin; anexcipient such as starch or lactose, a disintegrating agent such asalginic acid, Primogel, or corn starch; a lubricant such as magnesiumstearate or Sterotes; a glidant such as colloidal silicon dioxide; asweetening agent such as sucrose or saccharin; or a flavoring agent suchas peppermint, methyl salicylate, or orange flavoring.

When the dosage unit form is a capsule, it can contain, in addition tomaterial of the above type, a liquid carrier such as a fatty oil. Inaddition, dosage unit forms can contain various other materials whichmodify the physical form of the dosage unit, for example, coatings ofsugar, shellac, or other enteric agents.

The compound or its salts can be administered as a component of anelixir, suspension, syrup, wafer, chewing gum or the like. A syrup maycontain, in addition to the active compounds, sucrose as a sweeteningagent and certain preservatives, dyes and colorings and flavors.

In a preferred embodiment, the active compounds are prepared withcarriers that will protect the compound against rapid elimination fromthe body, such as a controlled release formulation, including implantsand microencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) are also preferred as pharmaceuticallyacceptable carriers. These may be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811 (which is incorporated herein by reference in its entirety).For example, liposome formulations may be prepared by dissolvingappropriate lipid(s) (such as stearoyl phosphatidyl ethanolamine,stearoyl phosphatidyl choline, arachadoyl phosphatidyl choline, andcholesterol) in an inorganic solvent that is then evaporated, leavingbehind a thin film of dried lipid on the surface of the container. Anaqueous solution of the compound is then introduced into the container.The container is then swirled by hand to free lipid material from thesides of the container and to disperse lipid aggregates, thereby formingthe liposomal suspension.

Suitable vehicles or carriers for topical application can be prepared byconventional techniques, such as lotions, suspensions, ointments,creams, gels, tinctures, sprays, powders, pastes, slow-releasetransdermal patches, suppositories for application to rectal, vaginal,nasal or oral mucosa. In addition to the other materials listed abovefor systemic administration, thickening agents, emollients, andstabilizers can be used to prepare topical compositions. Examples ofthickening agents include petrolatum, beeswax, xanthan gum, orpolyethylene, humectants such as sorbitol, emollients such as mineraloil, lanolin and its derivatives, or squalene.

Combination Treatment

The compound can also be mixed with other active materials which do notimpair the desired action, or with materials that supplement the desiredaction. The active compounds can be administered in conjunction, i.e.combination or alternation, with other medications used in the treatmentof disorders that are mediated by kinases or HSP90.

The compounds can be administered in combination or alternation withdrugs typically useful for treatment or prevention of asthma, such ascertain anti-inflammatory drugs and bronchodilators. Corticosteroids(inhaled and oral), mast cell stabilizers, and the leukotriene modifierdrugs are typically a useful anti-inflammatory medication for peoplesuffering from asthma. These drugs reduce swelling and mucus productionin the airways. Bronchodilators typically relieve the symptoms of asthmaby relaxing the muscle bands that tighten around the airways. Thisaction rapidly opens the airways, letting more air come in and out ofthe lungs. Bronchodilators also help clear mucus from the lungs.

Typically used compounds include Inhaled corticosteroids, which preventrather than relieve symptoms. Inhaled corticosteroids include: Advair (acombination medication that includes a corticosteroid (fluticasone) plusa long acting bronchodilator drug (in this case a β-2 adrenergicreceptor agonist, salmeterol)), aerobid (flunisolide), azmacort(triamcinolone), flovent (fluticasone), methylprednisolone, prednisone,pulmicort or serevent diskus (salmeterol powder), theophylline, qvar,and xopenex (levalbuterol), Inhaled corticosteroids come in three forms:the metered dose inhaler (MDI), dry powder inhaler (DPI) and nebulizersolutions. Systemic steroids include: methylprednisolone (Medrol,Methylpred, Solu-Medrol), prednisone (Deltasone) and prednisolone(Prelone, Pediapred, Orapred). Mast Cell Stabilizers include Intal andTilade, which work by preventing the release of irritating andinflammatory substances from mast cells. Leukotriene modifiers includeaccolate and singular and accolate (zafirlukast), singulair(montelukast) and zyflo (zileuton).

The compounds can be administered in combination with nonsteroidalantiinflammatories such as ibuprofen, indomethacin, fenoprofen,mefenamic acid, flufenamic acid, sulindac. The compound can also beadministered with corticosteriods. Any of the compounds described hereinfor combination or alternation therapy can be administered as anyprodrug that upon administration to the recipient, is capable ofproviding directly or indirectly, the parent compound. Nonlimitingexamples are the pharmaceutically acceptable salts (alternativelyreferred to as “physiologically acceptable salts”), and a compound whichhas been alkylated or acylated at an appropriate position. Themodifications can affect the biological activity of the compound, insome cases increasing the activity over the parent compound.

Processes for the Preparation of the Compounds

Modular synthetic processes directed to the synthesis of pochonin D andpochonin A were adapted to the synthesis of a library of resorcylic acidlactones that extend beyond the natural resorcylides. The synthesesdeveloped utilize resin-assisted or solid phase synthesis to minimizeand facilitate the isolation of intermediate and final products. First,a description of the synthetic protocols directed to the naturalresorsylic acid lactones is presented followed by the synthesis of thelibrary of compounds.

The following abbreviations are used herein.

Ac Acetyl (CH3C═O) ADP Adenosine diphosphate AIBNAzobis(isobutyronitrile) All Allyl ATP Adenosine triphosphate BERBorohydride exchange resin BBN Borabicyclononane Bn Benzyl Bz BenzoylCAN Ceric ammonium nitrate CSA Camphorsulfonic acid δ Chemical shift(NMR) dba Dibenzylideneacetone DBU 1,8-Diazabicyclo[5.4.0]undec-7-eneDDQ 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone DEAD Diethylazodicarboxylate DIAD Diisopropyl azodicarboxylate d.e.Diastereoisomeric excess DET Diethyl tartrate DHP Dihydropyran DIBAL orDiisobutylaluminum hydride Dibal-H DIC N,N′-diisopropylcarbodiimide DMAP4-Dimethylaminopyridine DMDO Dimethyldioxirane DMF DimethylformamideDMPI Dess-Martin periodinane DMSO Dimethylsulfoxide DNA Desoxyribonucleic acid dppe 1,2-Bis(diphenylphosphino)ethane EC₅₀ Plasmaconcentration required for obtaining 50% of maximum effect in vivo EDC1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride EDTAEthylenediaminetetraacetic acid e.e. Enantiomeric excess EOMEthoxymethyl (CH₃CH₂OCH₂—) FDA Food and Drug Administration Fmoc9-Fluorenylmethoxycarbonyl GI₅₀ Concentration required for 50%inhibition of cell growth Grubbs' II Grubbs' second generation catalyst:(ruthenium[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolinylidene)dichloro(phenyl-methylene)(tricyclohexylphosphane)

HFIP Hexafluoroisopropanol HMDS Hexamethyldisilazide HMPAHexamethylphosphorictriamide HOBT N-Hydroxybenzotriazole RCMRing-closing metathesis RedAl Sodium bis(methoxyethoxy) aluminum hydrideR_(f) Retention factor RNA Ribonucleic acid RT Room temperature SAESharpless asymmetric epoxidation SAR Structure-activity relationship SEM2-Trimethylsilylethoxymethoxy TBAF Tetra-n-butylammonium fluoride TBAITetra-n-butylammonium iodide TBDPS t-Butyldiphenylsilyl TBHPt-Butylhydroperoxide TBS t-Butyldimethylsilyl Teoc2-(Trimethylsilyl)ethoxycarbonyl Tf Triflate (CF₃SO₃) TFATrifluoroacetic acid TFAA Trifluoroacetic acetic anhydride THFTetrahydrofuran THP Tetrahydropyran TLC Thin layer chromatography TMSTrimethylsilyl Ts Tosyl (p-CH₃C₆H₄SO₂) p-TSOH para-Toluenesulfonic acid

I. Synthesis of Pochonin D

Preliminary Studies

Retrosynthetic disconnections for pochonin D (2-85) depicting thesynthetic strategy are shown below. A Mitsunobu esterification, anacylation and a ring-closing metathesis are shown as the maindisconnections using three building blocks: acid 2-87, alcohol (S)-2-27and Weinreb amide 2-88.

HPLC High performance chromatography HRMS High resolution massspectrometry HSP90 Heat shock protein 90 Hunig's BaseDiisopropylethylamine IC₅₀ Concentration of a drug that is required for50% inhibition in vitro imid. Imidazole Ipc₂BHBis-isopinocamphorylborane J Coupling constant KHMDS Potassiumhexamethyldisilylamide L.C. Liquid chromatography LDA Lithiumdiisopropylamide LiHMDS Lithium hexamethyldisilazide (LiN(SiMe₃)₂) μMMicromolar concentration (μmol · l⁻¹) MAP Mitogen-activated proteinmCPBA meta-Chloroperoxybenzoic acid MOM Methoxymethyl (CH₃OCH₂—) mRNAMessenger ribonucleic acid M.S. Mass spectrum NaHMDS Sodiumhexamethyldisilazide NMR Nuclear magnetic resonance NMMN-Methylmorpholine NMO N-Methylmorpholine-N-oxide NOE(SY) Nuclearoverhauser effect PCC Pyridinium chlorochromate PDC Pyridiniumdichromate PG Protecting Group PMB para-Methoxybenzyl PNA Peptidenucleic acid Piv Pivaloyl PS- Polymer supported PS-TBD(1,5,7)-Triaza-bicyclo[4.4.0]dodeca-5- ene-7-methyl polystyrene Pyr orPy Pyridine rac Racemic RAL Resorcylic acid lactone

Retrosynthetic analysis for pochonin D based on radicol and pochonin Csyntheses

The Weinreb amide moiety 2-88 was synthesized as shown in Scheme 1.Thus, alkylation of intermediate 2-7 with 5-iodo-1-pentene yieldedWeinreb amide 2-88 in two steps from thiophenol.

In parallel, an alternative synthetic pathway was developed, startingwith commercially available cis-6-nonen-1-ol (Scheme 2).

Following a classical oxidation procedure (PDC in DMF), acid 2-89 wasthen α-chlorinated by formation of the enolate using LDA and subsequentchlorine addition using carbon tetrachloride (Snider, B. B. & Kulkarni,Y. S., J Org Chem 1987, 52, 307-310). After work-up, compound 2-90 wasobtained as a black oil although as a pure compound by ¹H NMR. Effortsto purify this acid proved disappointed and it was used directly in thefollowing step. Further amide formation using N,O-dimethylhydroxylamineand EDC and displacement of the chlorine atom with thiophenol affordedcompound 2-92 in 74% overall yield from cis-6-nonen-1-ol. Via theoxidation/elimination reaction, the thioether Weinreb amide 2-92 couldbe converted in its closely related derivative 2-93.

Due to the high cost of commercially available2,4-dihydroxy-6-methylbenzoic acid, a protocol was developed that allowsthe synthesis of 2,4-dihydroxy-6-methylbenzaldehyde which can further bederivatized in the corresponding acid using various protecting groups(Scheme 3). Starting from orcinol and following a Vilsmeier-Haackprocedure, aldehyde 2-94a was obtained in 45% yield (72% based onrecovered S.M.). As described in the experimental section, this aldehydeprecipitates at pH=7 and is recovered with good purity (>95% as judgedby ¹H NMR).

Compound 2-94a was then protected with different groups to generatealdehydes 2-94c-k which were subsequently oxidized to afford thecorresponding acids 2-95a-k in good yields (Scheme 3, Table 2). Byvarying the oxidation conditions, it was found possible to chlorinatethe ring in a one pot sequence (oxidation/chlorination) in the requiredposition for the synthesis of pochonin D.

TABLE 2 Oxidation conditions to yield various protected resorcylic acidSolvent Yield 2-95 PG₁ PG₂ X Acid NaClO₂ sytem^(a) Time^(b) [%] a H H HNH₂SO₃H 3.25 equiv. A 1 h 86 b H H Cl NH₂SO₃H^(c) 2.0 equiv. B 12 h 90 cMe Me H NH₂SO₃H 3.25 equiv. A 1 h 82 d Me Me Cl NH₂SO₃H 3.25 equiv. B 12h 89 e EOM EOM H NaH₂PO₄  5.0 equiv. C 12 h 68 f EOM EOM Cl NaH₂PO₄  5.0equiv. B 12 h 89 g^(d) EOM H H NaH₂PO₄  5.0 equiv. C 12 h 87 h SEM H ClNH₂SO₃H 3.25 equiv. B 12 h 70 i TBDPS H Cl NH₂SO₃H 3.25 equiv. B 30 min93 j TBDPS TBDPS H NH₂SO₃H 3.25 equiv. A 30 min 92 k TBDPS TBDPS ClNH₂SO₃H 3.25 equiv. B 30 min 95 ^(a)A: H₂O/THF/DMSO (20:10:1) or B:H₂O/THF (2:1) or C: DMSO, ^(b)When run for more than 1 h, the reactionwas heated up slowly to room temperature, ^(c)In this case, only 2equiv. of acid were used to avoid over-chlorination, ^(d)This compoundwas previously assigned as compound 2-66.

Depending on the protecting group, two different acidic buffers (NH₂SO₃Hor NaH₂PO₄) and various solvent systems were used (Table 2). To avoidany chlorination reaction, a small percentage of DMSO as a mixture inTHF/H₂O along with sulfamic acid ((a) Lindgren, B. O. & Nilsson, ActaChem. Scand. 27, 888-890 (1973), (b) Colombo, L et al., J. Chem. Soc.,Perkin Trans. 1, 136-140 (1980)). proved to be essential and veryeffective to quench HOCl (entries a, c, j), (Dalcanale, E. & Montanari,F. Selective oxidation of aldehydes to carboxylic acids with sodiumchlorite-hydrogen peroxide. J Org Chem 51, 567-569 (1986)). The one potoxidation/chlorination sequence was done in absence of DMSO and requiredlonger time to reach completion (entries b, d, f, h). Due to the acidicliability of the EOM protecting groups, NaH₂PO₄ was used as instead ofsulfamic acid and the oxidation was carried out in pure DMSO. Thebis-protected compound was selectively deprotected to afford thechlorinated analog of compound 2-95g in 77% yield over the two steps(vide infra). In the first sequence used for the synthesis of thispochonin, the mono-MOM protected acid 2-96 was used. Mitsunobuesterification under standard conditions (DIAD, PPh₃) between this acid2-96 and racemic alcohol 2-27 afforded the desired ester 2-97 which wasfurther converted to the bis-protected ester 2-98 (Scheme 4).

Acylation using previously optimised conditions (2 equiv. of LDA at −78°C.) allowed the formation of the acyclic precursor 2-99 along with someunreacted starting material. Two different sequences(oxidation/elimination followed by ring-closing metathesis (path A) orvice-versa (path B)) to yield compound 2-102 were envisioned. Bothpathways proved similar (same overall yield) albeit with a higherselectivity in the metathesis reaction when following path B (cis/transratio 7:1 vs. 4:1 for path A). Having compound 2-102, MOM deprotectionand chlorination of the aromatic ring, was planned for the formation ofpochonin D (2-85, Scheme 5).

Deprotection of the MOM groups following a well-known procedure allowedeasily the generation of racemic monocillin II (2-103) but thechlorination to form pochonin D proved problematic.

As an alternative, starting with the chlorinated analog of acid 2-96 andfollowing the two steps sequence developed for MOM-protected MonocillinII (Scheme 4), compound 2-104 was obtained in good quantities (Scheme6). Although acylation reaction using Weinreb amide 2-7 led to theisolation of compound 2-103 in 37% yield, no reaction was observed whenusing Weinreb amide 2-88. As an alternate, the α,β-conjugated analog2-93 was used directly.

Indeed, acylation reaction between the toluic anion of ester 2-104 andWeinreb amide 2-93 led to the isolation of acyclic compound 2-107 in 40%yield along with some unreacted starting material. The possibility ofachieving the acylation reaction on the chlorinated toluic ester openedthe door for the completion of pochonin D synthesis.

Total Synthesis of Pochonin D

EOM protecting groups were chosen for phenolic protection. Using thechemistry described above to synthesize toluic acids, mono-EOMchlorinated acid 2-108 was synthesized in three steps from formylatedorcinol 2-94a (Scheme 7). The selective deprotection of the ortho-phenolwas achieved using a specific concentration of TFA in a THF/MeOH mixture(THF/TFA/MeOH 7:1.5:1 (vol.)) without any bis-deprotection. FurtherMitsunobu esterification using standard protocol (DIAD, PPh₃, toluene)and reprotection of the ortho-phenol allowed the formation of compound2-110 in 51% yield from acid 2-108.

Deprotonation of the toluic ester 2-110 followed by addition of Weinrebamide 2-93 afforded the desired metathesis precursor 2-111 in 52% yieldalong with some unreacted starting material. Treatment of triene 2-111with the Grubbs' second generation catalyst ((a) Chatterjee, A. K.,Morgan, J. P., Scholl, M. & Grubbs, R. H., J Am Chem Soc 122, 3783-3784(2000), (b) Scholl, M., Ding, S., Lee, C. W. & Grubbs, R. H., Org Lett1, 953-956 (1999)). at 120° C. for 15 min afforded the desiredcyclization product 2-112 in an excellent yield albeit as an unseparablemixture of cis/trans olefins 1:4. Metathesis reaction underthermodynamic control at 80° C. overnight (Lee, C. W. & Grubbs, R. H.Stereoselectivity of Macrocyclic Ring-Closing Olefin Metathesis. OrgLett 2, 2145-2147 (2000)) shifted the equilibrium to the transintermediate 2-112 with >95% selectivity (as judged by ¹H NMR) and 94%yield. It should be noted that this reaction could be performed atmillimolar concentration without any detectable amount of dimerizationor oligomerization. Importantly, the 10-membered ring macrocycle was notobserved. Further EOM deprotection using TFA in dichloromethane allowedthe first total synthesis of pochonin D which was found to have anidentical NMR spectrum to the natural product.

For the purpose of diversity-oriented synthesis and as the presence of athioether linkage was not possible having the chlorine atom on thearomatic ring, a more concise synthesis of pochonin D withpolymer-supported reagents was developed ((a) Ley, S. V. & Baxendale, I.R., Nat Rev Drug Discov 1, 573-86 (2002), (b) Ley, S. V. et al., J.Chem. Soc., Perkin Trans. 1 23, 3815-4195 (2000)). The Mitsunobureaction using directly 2,4-dihydroxy-6-methylbenzoic acid was envisagedleading to an even more concise pathway to pochonin D. As(S)-4-penten-2-ol ((S)-2-27) is commercially available, improvements tothe sequence leading to the aliphatic Weinreb amide 2-93 wereundertaken. A protocol using solid-support was developed to minimize theneed to purification of intermediates and final product. Commerciallyavailable 2-chloro-N-methoxy-N-methylacetamide (Scheme 8) wasselectively S-alkylated with 3-mercaptophenol using one equivalent ofbase, and then loaded onto Merrifield resin in the same pot reaction bythe successive addition of a second equivalent of K₂CO₃, the resin, andraising the temperature to 50° C.

This method afforded the polymer-bound Weinreb amide 2-49 in one step.Oxidation of the thioether 2-49 to the corresponding sulfoxide 2-113 wascarried out using the aforementioned procedure involving H₂O₂ inHFIP/CH₂Cl₂ (Ravikumar, K. S., Begue, J.-P. & Bonnet-Delpon, D. Aselective conversion of sulfide to sulfoxide in hexafluoro-2-propanol.Tetrahedron Letters 39, 3141-3144 (1998)). This oxidation procedure wasfound practical and reliable with no over-oxidation to the sulfone andeasy recycling of the fluorinated solvent as the reaction was carriedout on solid phase. The amidosulfoxide 99 was then deprotonated withtBuOK, and the resulting enolate was quenched with 5-iodo-1-pentene. Useof DMSO avoided sulfoxide elimination, and the reaction could be heatedup to 60° C. Resuspension of the resin in toluene and heating up to 80°C. released, after elimination, the desired fragment 2-114 with 77%yield and 95% purity (judged by ¹H NMR). This methodology on solidsupport was found very practical in terms of yield and purity of thebuilding block 2-114 as no column chromatography was needed.

Based on the chemistry developed in solution (Scheme 7), a selectiveMitsunobu esterification of 2,4-dihydroxy-6-methylbenzoic acid (2-95a)with (S)-4-penten-2-ol ((S)-2-27) using polymer-bound DEAD affordedester 2-116 (Scheme 9). The use of (mClPh)₃P was found essential tosuppress any competing ether formation with the para-phenol. Protectionof both phenols with EOM groups afforded non-chlorinated ester 2-117,which could be used in the subsequent alkylation without furtherpurification.

The chlorine was introduced prior to esterification using HClO generatedin situ by the oxidation of acetaldehyde with NaClO₂/sulfamic acid. Acid2-95b was obtained from its non-chlorinated parent 2-95a in 92% yieldwithout seeing any over-chlorination. Esterification of this productunder the same conditions as for 2-95a afforded compound 2-115. Furtherbis-protection with EOM groups led to ester 2-110 which could also beused in subsequent reactions without any purification. Deprotonation ofthe toluic esters 2-110 and 2-117 followed by addition of Weinreb amide2-114 afforded the desired metathesis precursors 2-118 and 2-119 whichcould be used directly in the following step (Scheme 10). When theacylation reaction was performed on the non-chlorinated ester 2-117, 20%of product stemming from a 1,4-conjugated addition on the Weinreb amidewas obtained.

Treatment of crude trienes 2-118 and 2-119 with the Grubbs' secondgeneration catalyst under thermodynamic control at 80° C. overnight ledexclusively (more than 95% selectivity) to the trans macrocycles 2-112and 2-120. A simple filtration through a path of silica was then used toremove all of the catalyst and its by-products, affording compound 2-112in 44% yield over two steps. While the metathesis reaction carried outon purified triene 2-118 was nearly quantitative, it was found morepractical to carry out the whole synthetic sequence from compound 2-95awithout any purification, thus affording the protected pochonin D 2-112in 25% yield over five steps. Purification with column chromatographyisolated 2-120 from the 12-membered macrocycle 2-121. Removal of EOMgroups from both macrocycles 2-112 and 2-120 using sulfonic acid resinin MeOH allowed the synthesis of both pochonin D (2-85) and monocillinII (2-103) in 90% and 92% yield, respectively. As shown for theacylation reaction, the presence or absence of the chlorine atom on thearomatic ring seems to influence the reactivity of the conjugatedolefin. Indeed, deprotection of compound 2-120 with HCl (2.5% indioxane) led to the conjugated addition of the chlorine ion, whereascompound 2-112 could be deprotected with HCl to obtain pochonin D(2-85). This synthesis using polymer-supported reagents allowed theachievement of pochonin D and monocillin II in six (23% yield) and five(24% yield) steps respectively. Starting from commercially availablebuilding blocks, only one chromatographic purification of the finalcompound is required for all the synthetic pathway and this methodologycould be used for the synthesis of libraries. Evaluation for HSP90affinity in a competition assay with geldanamycin revealed that pochoninD is a good ligand for HSP90 with an IC₅₀ of 80 nM as compared to 20 nMfor radicicol (vide infra).

II Synthesis of Pochonin A

Having access to pochonin D (2-85), pochonin A (2-122) was then preparednot only to confirm its structure but also to compare its biologicalactivity to pochonin D and radicicol.

Epoxidation of pochonin D using DMDO allowed the formation of pochonin Aas a 1:1 mixture of diastereoisomers that could be separated by columnchromatography. An alternate route is depicted in Scheme 11. Epoxidationof bis-EOM protected pochonin D (2-112) also gave the epoxide as amixture of diastereomers.

The suitability of silyl based protecting groups to access this type ofmolecules was investigated. Thus, persilylation of benzoic acid 81b(Scheme 12), followed by “acid-free” conversion of the silyl ester tothe corresponding acyl chloride yielded key intermediate 112 uponesterification with alcohol (R)-27 (Wisnner, A. & Grudzinskas, C. V., JOrg Chem 43, 3972-3974 (1978)).

Deprotonation of the toluic ester 2-126 followed by reaction withWeinreb amide 2-114 afforded metathesis precursor 2-127. Ring-closingmetathesis using Grubbs' second generation catalyst under aforementionedthermodynamic conditions (80° C., overnight) afforded macrocycle 2-128in good yield and excellent cis/trans ratio (<5% cis). Epoxidation ofthe non-conjugated olefin was optimal when carried out at 0° C. withmethyl(trifluoromethyl)-dioxirane generated in situ (Yang, D., et al. J.Org. Chem., 60, 3887-3889 (1995) affording TBS-protected pochonin A(2-129) in excellent yield as an inseparable 3:1 diastereoisomericmixture. Deprotection of compound 2-129 using classical silyldeprotection conditions (TBAF in THF) afforded a separablediastereomeric mixture and confirmed that the major product was indeedthe desired pochonin A (2-122).

Alternative protecting groups were evaluated to improve the yield of theesterification and acylation reactions. Based on their stability towardbasic conditions but also on their liability towards TBAF, SEMprotecting groups were considered. Following the procedure described forthe polymer-assisted synthesis of pochonin D, selective Mitsunobureaction between benzoic acid 2-95b and chiral alcohol (S)-2-27 usingpolymer-bound DEAD and subsequent protection with SEM-Cl afforded ester2-130 in 72% yield (Scheme 13).

Acylation of toluate ester 2-130 using Weinreb amide 2-114 led to theisolation of the acyclic precursor 2-131 in 60% yield. Treatment of thetriene 2-131 with the Grubbs' second generation catalyst underthermodynamic conditions (80° C., overnight) afforded the correspondingmacrocycle 2-132 in 87% yield (<5% cis olefin), which was epoxidizedunder the same conditions as for the TBS-protected compound 2-128[methyl(trifluoromethyl)-dioxirane], yielding compound 2-133 in 83%yield albeit in a 1:1 diastereomeric ratio (inseparable). Deprotectionof 2-133 was achieved with 8.0 equiv. of MgBr₂.Et₂O affording thedesired pochonin A (2-122) along with its diastereoisomer (2-122b) as aseparable mixture. Pochonin A was found to be a good ligand of HSP90with an IC₅₀ of 90 nM (vide infra).

III. Diversity Oriented Synthesis of Pochonin Analogues

With the objective to extend the diversity of the compounds in the hopeof finding new ATPase and kinase inhibitors, a library of pochoninanalogues was prepared and evaluated for biological activity. Thislibrary was envisioned to stem from five points of diversity around theresorcylic macrolide scaffold: modifications of the para-phenol (R¹, anumber of natural resorcylides bearing a methyl group at this position),the group on C17 (R², both stereochemistry are present in naturalresorcylides; however, only with a methyl substituent),

the C14-C15 olefin (R³), the C9 carbonyl (R⁴), the olefin C10-C11, andthe meta position on the aryl ring (R⁵, a number of natural resorcylidesbear a chlorine at this position). To minimize traditionalchromatography, the synthesis was conducted with polymer-bound reagents.The assembly of the macrocycle relied on the chemistry developed for thesynthesis of pochonin D using polymer-bound reagents (Schemes 9 and 10).

A variety of homoallylic alcohols 2-134 bearing various substituent atR² was prepared. Homoallylic alcohols 2-134 that are not commerciallyavailable may be obtained in by any suitable method. In one embodiment,the homoallylic alcohols 2-134 were obtained in their highestenantiomeric form either by enzymatic resolution of the racemic alcohol(H. E. Master et al., Tet. Lett., 37:9253 (1996); S. Singh et al., Tet.Asymm., 13:2679 (2002) or via Brown allylation of the correspondingaldehyde (H. C. Brown and P. K. Jadhav J. Am. Chem. Soc., 105:2092(1983). The phenyl (2-134a), the pyridinyl (2-134b) and the furyl(2-134c) alcohols were prepared by enzymatic resolution (Scheme 14).Racemic alcohols 2-134a-c were obtained after Grignard addition ofcommercially available allylmagnesium bromide on their correspondingaldehyde 2-134a-c.

Kinetic enzymatic resolution of racemic alcohols 2-134a-c was realizedusing the highly efficient Amano lipase (an immobilized version ofPseudomonas cepacia). This enzyme catalyzed a selectivetransesterification of alcohols (R)-2-134a-c with vinyl acetate as anacyl donor, the (S) alcohols 2-134a-c being isolated in excellent yieldsand good enantiomeric excesses (Table 3).

TABLE 3 Enantioselective acylation of alcohols rac-2-134a-c bytransesterification with lipase Conv. Yield e.e. Yield e.e. Time Ratio(%) (%) (%) (%) (%) Entry Substrate (h) (OH/OAc) (S)-2-134 (S)-2-134(R)-2-134 (R)-2-134 1 rac-2-134a 30 50:50 45 98 49 93 2 rac-2-134b 3052:48 50 89 39 94 3 rac-2-134c 5 49:51 44 88 49 89

Enantiomeric excess obtained with this methodology are all above 88%.Acetylated alcohols (R)-2-137 were then hydrolysed to the correspondingalcohols (R)-2-134a-c in excellent yields. In addition, a second processbased on Brown allylation was developed for the synthesis of theisopropyl (2-134d), the propyl (2-134e) and the benzyl (2-134f) alcohols(Scheme 15).

(−)-B-Allyldiisopinocampheylborane (2-139, (−)-Ipc₂BAllyl) wassynthesized in a three steps sequence from (−)-α-pinene involving anhydroboration, the formation of the corresponding MeO-borinic ester2-138 and its treatment with a Grignard reagent. Further condensation onaldehydes 2-134d-f followed by oxidation of the resulting borinates withalkaline hydrogen peroxide allowed the formation of the chiralhomoallylic alcohols 2-134d-f in good enantiomeric excess.

The macrocycle assembly was modeled after the synthesis of pochonin D.(E. Moulin, V. Zoete, S. Barluenga, M. Karplus, N. Winssinger, J. Am.Chem. Soc., 127:6999 (2005)). Thus as shown in Scheme 16, commerciallyavailable benzoic acid 2-95a and its chlorinated analog 2-95b (thechlorine atom was introduced on acid 2-95a prior to esterification usingHClO generated in situ by the oxidation of acetaldehyde withNaClO₂/sulfamic acid were esterified with a variety of homoallylicalcohols using polymer-supported DEAD to yield esters 2-115a-g and2-116a-g in excellent purity. (E. Moulin et al., J. Am. Chem. Soc.,127:6999 (2005)). The products 2-115a-g and 2-116a-g were then protectedwith ethoxymethylene chloride (EOM-Cl) in the presence of Hunig's baseto obtain the corresponding protected toluic esters 2-110a-g and2-117a-g which could be used in the subsequent carbon-acylation reactionwithout further purification.

Deprotonation of the toluic esters 2-110a-g and 2-117a-g using twoequivalents of LDA, followed by addition of the α,β-unsaturated Weinrebamide prepared via solid phase chemistry (E. Moulin et al., J. Am. Chem.Soc., 127:6999 (2005)) afforded acylation products 2-118a-g and2-119a-g. The reaction was quenched with a polymer bound acid which alsosequestered all the diisopropyl amine. This reaction can lead to somelevel of 1,4-conjugate addition product (S. Barluenga et al., Chem. Eur.J., 11:4935 (2005)). While the bulky chlorine present in pochonin Dsuppresses this reaction, compounds lacking the aryl chloride afforded20% of the conjugate addition products 2-140a-g. Nevertheless, the crudemixtures of these reactions were used in the subsequent cyclizationstep. The trienes were then subjected to ring closing metathesis usingGrubbs' second generation catalyst (A. K. Chatterjee et al., J. Am.Chem. Soc., 122:3783 (2000); M. Scholl et al., Org. Lett., 1:953 (1999))under thermodynamic conditions (C. W. Lee, and R. H. Grubbs, Org. Lett.,2:2145 (2000)), affording the desired 14-membered macrocycles. In thecases where metathesis reactions were carried out with a mixture of2-118a-g, 2-119a-g and 2-140a-g, the corresponding 12-membered ringproducts 2-121a-g were obtained in addition to 2-112a-g and 2-120a-g asa separable mixture. All successful reaction sequences were purified bystandard chromatography yielding the macrocycle 2-112a-g and 2-120a-gand 2-121a-g in 30-60% and 8-10% overall yield respectively from 2-95.

Macrocycles 2-112a-g and 2-120a-g were then used as the starting pointfor further diversifications. Deprotection of the EOM groups of 2-112a-gand 2-120a-g using sulfonic acid resin afforded compounds 2-103a-g and2-85a-g in pure form and excellent yields after simple filtration of theresin and evaporation of the solvents (Scheme 17). The 12-membered ringproducts 2-121a-g were deprotected just as smoothly (not shown).Treatment of 2-112a-g and 2-120a-g with reducing agents led to eithercarbonyl reduction using Dibal or mixtures of carbonyl and 1,4-reductionwith NaBH₄. It is known that using non-coordinating counter ion forborohydride can favor the carbonyl reduction (H. W. Gibson and F. C.Baily, J. Chem. Soc. Chem. Commun., 1977:815; A. Kirschning, J. Prakt.Chem., 2000:342). This was most conveniently achieved using a polymersupported quaternary ammonium borohydride known as borohydride exchangeresin (BER). Thus, ketones 2-112a-g and 2-120a-g could be reduced usingBER-resin to obtain both diastereoisomers of 2-141a-g in ˜60% yield.Deprotection of the EOMs with sulfonic acid resin under regularconditions afforded compounds 2-142a-g. Acetylation of the reducedintermediates 2-141a-g using PS-NMM/Ac₂O yielded compounds 2-143a-gwhich led to elimination upon deprotection to afford trienes 2-144 as amixture of olefin geometries.

Prolonged exposure of resorcylides 2-112a-g and 2-120a-g to methanol inthe presence of sulfonic acid resin was found to lead to conjugateaddition; this observation was exploited to drive the reaction tocompletion cleanly. Thus phenol 2-85a-g was quantitatively converted toproduct 2-145a-g in 15 h (Scheme 18). This product could obviously beobtained directly from 2-120a-g under the same conditions.

Compounds 2-103a-g and 2-85a-g were also used as the starting points forfurther diversifications (Scheme 19). Thus, treatment of 2-103a-g and2-85a-g with polymer-bound cyanoborohydride afforded the 1,4-reductionproducts 2-146 in moderate yields. The more acidic para-hydroxyl groupsof 2-103a-g and 2-85a-g were substituted via either Mitsunobu reactionusing polymer bound DEAD or alkylation using a polymer-bound base toafford compounds with general structure 2-147 and 2-148 respectively.Oxidation with OsO₄ afforded the dihydroxylation products 2-149 as amixture of isomers as well as the products corresponding to thedihydroxylation of the conjugate olefin (product not shown). Treatmentof 2-103a-g and 2-85a-g with freshly prepared dimethyldioxirane led tothe selective epoxidation of the non-conjugated olefin as a mixture ofdistereoisomers of pochonin A analogues 2-150. Although higherdiastereoselectivity may be obtained for pochonin A if the phenols areprotected with TBS (E. Moulin et al., Org. Lett., 7:5637 (2005)),protection was not needed here.

It is interesting to note that the conjugated olefin proved to havedifferent reactivity depending on the presence or absence of thechlorine atom on the aryl ring. Whereas EOM deprotection of compound2-120 where X=Cl and R=Me could be carried out with HCl in dioxane,treatment of the corresponding compound 2-112 where X═H and R=Me by thesame conditions led to the conjugate addition of the chlorine ion duringthe deprotection, affording compound 2-151 (Scheme 20). The β-chlorinecould be cleanly eliminated in the presence of polymer-bound base torecover the conjugate compound 2-103a-g.

While evaluating protecting groups for the phenols, it was noticed thatdihydropyran, in the presence of a strong acid such as sulfonic acid,led to electrophilic aromatic substitution rather than phenolprotection. (see also T. Kometani et al., Synthesis, 1988:1005).Applying these conditions to compounds 2-103 (Scheme 20) afforded 2-153as a separable mixture of diastereoisomers.

Compounds 2-120 (X=H, Scheme 20) protected with EOM groups underwentsmooth oxime formation with nine different hydroxylamines to obtaincompounds 2-154 as E/Z mixtures with variable ratios. EOM deprotectionof 2-154 with sulfonic acid resin in methanol followed by treatment withsulfonic acid resin in DCM in the presence of dihydropyran affordedoximes 2-155 bearing a pyran substitution on the aromatic ring as amixture of diastereoisomers. In the case where the side chain containsan acid (R²X═OCH₂COOH), the deprotection of the EOM with the sulfonicacid resin in methanol was accompanied by esterification of thecarboxylate. Treatment of compounds 2-120a and 2-112a withtrifluoroacetic acid led to the formation of trifluoroacetate 2-152.Oxime formation in the chlorinated analogues with or without EOMprotecting groups generated mostly the corresponding 1,4-addition of thehydroxylamines (Scheme 21). Surprisingly, when pochonin D was protectedwith TBS groups, (2-128a-g, Scheme 21) the formation of the desiredoxime 2-157a-g was the only product observed under the same reactionconditions. Deprotection of the TBS groups was than achieved using TBAFto obtain oximes 2-158a-g. Treatment of pochonin D with hydroxylaminesled to the formation of the 1,4-addition product 2-156.

In another embodiment, the bis-methylated compounds 2-164 were prepared(Scheme 22). Acid 2-108 was used in a standard Mitsunobu esterificationwith alcohol 2-159. Compound 2-160 was protected as the ortho-phenolfollowed by acylation reaction with Weinreb amide 2-114 to yield theacyclic precursor 2-162. Ring-closing metathesis followed by removal ofthe EOM protecting groups on compound 2-163 using sulfonic acid resinfurnished the bis-methylated analog 2-164.

Additionally, oxime derivatives 2-165 and 2-167 were synthesized frommacrocycle 2-163 as a separable mixture with the 1,4-addition product(Scheme 23). The carboxylic acid moiety of oxime 2-167 was thenesterified to form the corresponding piperidine amide oxime 2-168.Removal of the EOM groups using sulfonic acid resin allowed theisolation of both oximes 2-166 and 2-167 from 2-165 and 2-168respectively.

In another embodiment, the pochonin oximes were prepared fromcyclization of the pre-formed oximes as shown in Scheme 24. For exampleprotected pre-formed oxime 1 was bound to Wang resin usingN,N′-diisopropylcarbodiimide and dimethylaminopyridine. The allylprotecting group was removed and the phenol was functionalized underMitsunobu conditions. This was followed by deprotection of thecarboxylic acid and esterification with R³OH. Ring-closing was effectedwith the use of Grubbs II catalyst as described before to provideresin-bound oxime 5. Deprotection and removal from the resin wasachieved with trifluoroacetic acid to provide oxime 6. The carboxylicacid can be reacted with a variety of groups R⁴XH to provide the oximes7, wherein X is oxygen, sulfur, amino or substituted amino. The oximes 7were generally obtained as a 1:1 mixture of E:Z isomers which could beseparated by chromatography.

In another embodiment, the pochonin oximes were prepared by an alternateprocess which utilized a Mitsunobu esterification to construct themacrocylic ring. Scheme 25 shows a non-limiting example for theformation of the macrocycle 2-a1 via a Mitsunobu esterification. Orcinol(compound 8) is oxidized with phosphoryl chloride in DMF to providealdehyde 9, which is protected as the bis-ethoxymethyl ether andsubjected to oxidative chlorination with NaClO₂ to produce carboxylicacid 10. The carboxylic acid is protected as a trimethylsilylethyl esterand treated with LDA and Weinreb amide 12 (Scheme 26) to provide theα,β-unsaturated ketone 13. Ketone 13 is reacted withcarboxymethoxylamine hemihydrochloride in pyridine at 40° C. to producethe corresponding oximes as a mixture of E and Z isomers. The oximeswere converted to the desired amides 14 as a mixture of E and Z isomersby treatment with EDC and piperidine. Macrocyclization of precursor 14to protected compound 15 was performed under Mitsunobu conditions byslow addition of DIAD to a solution of compound 14 and PhP₃ in toluene.The phenol groups were deprotected by treatment with sulfonic acid resinat 40° C. to produce 2a-1 as a mixture of E and Z isomers. The mixtureof E- and Z-isomers was separated to obtain the pure E and Z isomers2a-1.

The diene Weireb amide 12 was prepared according to the process shown inScheme 26. Trans-3-hexenedioic acid dimethyl ester 16 was reduced to thecorresponding diol with lithium aluminum hydride. The diol wasmono-protected as the tert-butyldiphenylsilyl ether 17, and the freealcohol was converted to aldehyde 20 in three steps via the nitrile 19.Aldehyde 20 was then treated with Weireb amide ylide 21 to produce thediene Weireb amide 12, which was used to prepare compound 13 (Scheme24).

Crystal structures of the E-izomer 2a-1 and the related E-oxime 2a-13,which lacks the chloro substituent on the aromatic ring, were obtained.The crystal structures are shown in FIGS. 1 and 2.

The solubility of compounds 2a-1 and 2a-13 was determined. Bothcompounds were determined to be highly soluble in DMSO and DMA (>5g/mL). The good solubility of the compounds in DMSO and DMA enableformulations for intravenous or intraperitoneal administration. In onenon-limiting example, a formulation of the oximes in DMSO/Tween 20/0.9%NaCl (Oct. 5, 1985) was prepared.

Biological Activity

A representative subset of the pochonin analogue library (84 compounds)was tested for its inhibition in a panel of 24 kinase (AKT1, ARK5,Aurora-A, Aurora-B, B-RAF-VE, CDK2/CycA, CDK4/CycD1, CK2-α1, FAK, EPHB4,ERB2, EGF-R, IGF1-R, SRC, VEGF-R2, VEGF-R3, FLT3, INS-R, MET, PDGFR-β,PLK1, SAK, TIE2, COT) at 10 μM. A description of the assay method andresults is presented in Example 24.

Significantly, twelve compounds showed more than 50% inhibition, whichrepresents a >14% hit rate for a kinase. Surprisingly, pochonin D,pochonin A and radicicol, though they had been shown to be powerfulinhibitors of HSP90, showed no significant activity against this panelof kinases. Nine compounds were selected to calculate IC₅₀ against eachof the 24 kinases (table 4). In this more detailed analysis, radicicolshowed only very mild activity against VGFR-R2 with no inhibition forthe twenty-three other kinases. Several pochonin analogues showed awell-defined pattern of activity against therapeutically relevantenzymes such as Src (8 μM for A2), Aurora A (12 μM for A3), IGF1-R (11μM for A5). Importantly, the compounds that proved to be kinaseinhibitors were not inhibitors of HSP90 and are not indiscriminateATP-surrogates.

Another subset of the library was tested for HSP90 inhibition bymeasuring direct interaction in a competitive assay and measuringdepletion of HSP90 client proteins in a cellular assay. HSP90's ATPasepocket has a specific fold that is present in a superfamily whichincludes functionally diverse proteins such as DNA topoisomerase II,helicase, MutL and histidine kinases (Bergerat fold). (A. Bergerat etal., Nature, 386:414 (1997); R. Dutta and M. Inouye, Trends Biochem.Sci., 25:24 (2000)). In fact, it has been shown that radicicol doesinhibit other members of this family albeit with lower affinity. (D.Gadelle et al., Nucleic Acids Res., 33:2310 (2005); P. G. Besant et al.,Mol. Pharmacol., 62:289 (2002). Yet remarkably, the best HSP90inhibitors of the invention were selective for HSP90 with respect to thepanel of kinases. Sixteen compounds were found to have an IC50<1 μM.

Pochonin Analogs Selected for IC₅₀ Determination

TABLE 4 Inhibitory activity (IC₅₀: μM) of selected pochonin analogs in apanel of 24 kinase assays (a blank represents an IC₅₀ >50 μM). Radi-Resorcylic Acid Species Kinase cicol A-1 A-2 A-3 A-4 A-5 A-6 A-7 A-8AKT1 ARK5 Aurora-A 14 12 30 47 Aurora-B 16 14 36 16 B-RAF-VE 50CDK2/CycA CDK4/CycD1 50 30 48 45 37 CK2-α1 FAK 14 9 37 38 34 EPHB4 16 4049 40 ERBB2 16 24 EGF-R 33 10 14 22 16 32 IGF1-R 23 16 19 13 11 21 SRC11 8 14 12 12 20 VEGF-R2 49 19 20 30 19 25 VEGF-R3 40 19 31 31 34 FLT323 23 45 44 INS-R 36 44 MET 32 29 36 PDGFR-β PLK1 SAK 17 25 19 20 17TIE2 72 16 15 25 COT

The ATP-binding pocket of HSP90 targeted by radicicol and pochonin D hasa specific fold that is present in a superfamily which includesfunctionally diverse proteins, such as DNA topoisomerase II, helicase,MutL, and histidine kinases (R. Dutta and M. Inouye, Trends Biochem.Sci., 24:24 (2000)). In fact, it has been shown that radicicol doesinhibit other members of this family albeit with lower affinity (D.Gadelle et al., Nucleic Acids Res., 33:2310 (2005); P. G. Besant et al.,Mol. Pharmacol., 62:289 (2002)). While the pochonin library describedabove will certainly contain some compounds that are good inhibitors ofenzymes bearing a Bergerat fold, we wished to evaluate whethermodification around the pochonin scaffold could retune the selectivityof these compounds from HSP90 inhibitors to kinase inhibitors. The factthat more than fourteen percent of the compounds showed a kinaseinhibition of greater than 50% at 10 mM clearly supports the hypothesisthat RAL is a good scaffold for kinase inhibition.

Screening of the library for HSP90 affinity in the competition assay ledto the following general trends regarding structure activityrelationships (numbering and nomenclature based on formula A): Whilelarge substituents at R₁ are well tolerated for kinase activity, methylor hydrogen were generally found to be better for HSP90 affinity;substitution at R₂ generally led to a decrease of HSP90 affinity; alkeneat C14-15 (AB) were generally comparable in activity to thecorresponding epoxide for HSP90 affinity however the corresponding diolswere generally less active and trans alkenes were better than a cisalkenes; the presence of an alkene at C10-11 (CD) was generally found tobe superior to the corresponding alkane or the products of substitutionat C11 for HSP90 affinity; reduction of the C9 carbonyl generally led tolower HSP90 affinity while some substitution with hydroxyl aminesincrease affinity for HSP90; substitution at R₄ with small groups suchas chlorine was generally found to improve HSP90 activity except in somepermutations of substitutions at R₁ and R₃ where a hydrogen is superiorto a chlorine; substitutions of the phenols are tolerated of increaseaffinity for kinases but generally decrease affinity for HSP90.

It is important to note that generally, the compounds that showed potentaffinity for HSP90 were not good inhibitors of kinases stronglysuggesting that this scaffold is not an indiscriminate purine agonist.

Antitumor Activity

The compounds of the invention were evaluated for antitumor activity inmice. Tumors were induced intraperitoneally in healthy CB17/SCID mice byinjecting HCC1954 cells into the right flank of 70 mice. Once theinduced tumors were an average size of 80 mm, the mice were treated withcompound a2-1 at 25 mg/kg, 50 mg/kg and 100 mg/kg. Controls of vehiclealone and Herceptin® at 10 mg/kg were utilized. A dose-dependentreduction in tumor size was observed for the mice which were treatedwith compound 2a-1. The results were superior to those obtained in micetreated with Herceptin®, even at the 25 mg/kg dose for three weeks.Importantly, the re-growth of tumor after the last treatment did notoccur for three to seven days, indicating that a administration scheduleof one to three times per week should be effective.

Example 26 shows the antitumor activity of selected pochonin analogsagains HCC1954 and SK-BR-3 tumor cells. Compounds showing significantcytotoxixty were further examined for their ability to inducedegradation of known HSP90 client proteins such as ErbB2 in SK-BR3.Thus, after 18 hrs treatment with the compounds, the whole cell proteinlysates were obtained, protein concentrations were normalized and theconcentration of ErbB2 was quantified by Western blotting (C. Chavany etal J. Biol. Chem. 271:4974-4977 (1996)). In this assay compound 2a-1 hadan EC₅₀ of 26 nM and compound 2a-13 had an EC₅₀ of 12 nM whereasradicicol had an IC₅₀ of 289 nM. It is noteworthy that one oxime isomerwas systematically more active than the other. For 2a-11 and 2a-13, theE-isomer was 5 to 10 times more active than the Z-isomer.

Maximum Tolerated Dose

Three healthy CB17/SCID mice were treated with compound 2a-1 at 25mg/kg, 50 mg/kg and 100 mg/kg once daily for five consecutive days. Micetreated at the 25 mg/kg concentration did not show any weight loss. Micetreated at the 50 mg/kg and 100 mg/kg dose levels show a slight, butacceptable body weight loss.

EXAMPLES

General Techniques. All reactions were carried out under a nitrogenatmosphere with dry (anhydrous) solvents under anhydrous conditions,unless otherwise noted. Anhydrous solvents were obtained by passing themthrough commercially available alumina column (Innovative Technology,Inc.,® VA). All substituted polystyrene resins (100-200 mesh, 1% DVB)were purchased from Novabiochem® or Aldrich®. The Grubbs' II catalystwas purchased from Materia Inc.® Solid phase reactions were carried on aQuest® 210 or round bottom flasks and filtered in fritted funnels.Reactions were monitored by thin layer chromatography (TLC) carried outon 0.25 mm E. Merck® silica gel plates (60F-254) using UV light asvisualizing agent and 10% ethanolic phosphomolybdic acid or vanillinsolution and heat as developing agents. E. Merck® silica gel (60,particle size 0.040-0.063 mm) was used for flash column chromatography.PTLC (preparative thin layer chromatography) were carried out on 0.25 mmE. Merck® silica gel plates. NMR spectra were recorded on a BrukerAdvance-400® instrument and calibrated by using residual undeuteratedsolvent as an internal reference. The following abbreviations were usedto explain the multiplicities: s=singlet, d=doublet, t=triplet,q=quartet, m=multiplet, b=broad. IR spectra were recorded on aPerkin-Elmer 1600 series FT-IR spectrometer. LC-MS were recorded usingan Agilent 1100® HPLC with a Bruker® micro-TOF instrument (ESI). Unlessotherwise stated, a Supelco® C8 (5 cm×4.6 mm, 5 μm particles) column wasused with a linear elution gradient from 100% H₂O (0.5% HCO₂H) to 100%MeCN in 13 min at a flow rate of 0.5 ml/min. Unless otherwise stated,LDA was prepared at a concentration of 0.566 M by treating a solution ofdiisopropylamine (1.0 equiv.) in THF at −78° C. with n-butyllithium (1.0equiv.) and stirred for 30 min at this temperature before use.

Example 1 General Procedure for the Synthesis of Compounds 2-110

As depicted in Scheme 16, a solution of acid 2-95A or 2-95b (1.0 equiv),homoallylic alcohol (1.0 equiv) and tris-(3-chlorophenyl)phosphine (2.0equiv) in anhydrous toluene (0.05 M) was treated at room temperaturewith PS-DEAD (2.5 equiv, 1.3 mmol g⁻¹). After stirring for 10 min, thereaction mixture was filtered on silica and washed with hexane/EtOAc(10/1, 100 ml) and hexane/EtOAc (3/1, 100 ml). The 3/1 mixture wasconcentrated under reduced pressure to yield compound 2-115 (60-80%).Without further purification, compound 2-115 (1.0 equiv) andtetrabutylammonium iodide (catalytic amount) were dissolved in DMF (0.15M) and treated with diisopropylethylamine (4.0 equiv) and(chloromethyl)ethyl ether (4.0 equiv). After stirring overnight at 80°C., the reaction mixture was diluted with EtOAc and washed several timeswith a saturated NH₄Cl solution. The organic phase was dried over MgSO₄and concentrated under reduce pressure to yield compounds 2-110(80-90%). Using this method, a variety of compounds 2-110 were prepared.

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.04 (s, 1H), 5.89 (ddt, J=17.0, 10.5,7.0 Hz, 1H), 5.31 (s, 2H), 5.21 (s, 2H), 5.22-5.06 (m, 3H), 3.79 (q,J=7.0 Hz, 2H), 3.72 (q, J=7.0 Hz, 2H), 2.48-2.44 (m, 2H), 2.36 (s, 3H),2.01 (qd, J=12.4, 7.0 Hz, 1H), 1.25 (t, J=7.0 Hz, 3H), 1.23 (t, J=7.0Hz, 3H), 1.02 (d, J=6.4 Hz, 3H), 1.01 (d, J=7.0 Hz, 3H); ¹³C NMR (CDCl₃,100 MHz, 25° C.) δ 167.3, 154.0, 152.9, 134.8, 134.1, 120.4, 117.5,117.1, 101.5, 93.9, 93.4, 79.0, 64.6, 64.3, 35.6, 30.8, 18.4, 17.6,17.5, 15.0 (×2); HRMS (ESI-TOF) m/z 437.1574 ([M+Na⁺], C₂₁H₃₁O₆ClNarequires 437.1701).

The following non-limiting examples of compounds 2-217 were prepared.

Example 2 General Procedure for the Synthesis of Compounds 2-118, 2-119and 2-140

As depicted in Scheme 16, a solution of compound 2-110 or 2-117 (1.0equiv) in anhydrous THF (0.2 M) was treated at −78° C. with freshly madeLDA (2.0 equiv). Immediately after, the α,β-unsaturated Weinreb amide(S. V. Ley and I. R. Baxendale, Nat. Rev. Drug Discov., 1:573 (2002))was added to the cooled solution (1.0 equiv). The resulting mixture wasthen stirred for 10 min at −78° C. and quenched by addition ofAmberlite® resin (20 equiv). Upon warming up to room temperature, thereaction was filtered on a pad of silica and washed with EtOAc.Concentration under reduced pressure afforded the desired compound 2-118or 2-119. This compound was used directly in the metathesis reactionwithout any further purification. When X=H, 20% of the corresponding1,4-addition compound was observed and a fraction of the mixture waspurified for characterization of compounds 2-118/2-119 and 2-140 (SiO₂,0-20% EtOAc/cyclohexane gradient). Illustrative examples of compounds2-118/2-119 and 2-140 follow; their characterization followsrespectively in the text below each figure.

Illustrative Compound 2-119:

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 6.91 (dt, J=15.8, 6.7 Hz, 1H), 6.87(d, J=1.8 Hz, 1H), 6.53 (d, J=1.8 Hz, 1H), 6.19 (d, J=15.8 Hz, 1H),5.91-5.77 (m, 2H), 5.23 (s, 2H), 5.22 (s, 2H), 5.15-5.00 (m, 5H), 3.87(s, 2H), 3.73 (q, J=7.0 Hz, 4H), 2.45-2.40 (m, 2H), 2.35-2.29 (m, 2H),2.25-2.20 (m, 2H), 2.00-1.92 (m, 1H), 1.24 (t, J=7.0 Hz, 6H), 1.00 (d,J=2.3 Hz, 3H), 0.98 (d, J=3.0 Hz, 3H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.)δ 196.3, 167.6, 159.0, 156.2, 147.1, 137.0, 135.0, 134.4, 129.5, 118.7,117.2, 115.5, 111.0, 102.3, 93.3, 93.0, 78.8, 64.4, 64.3, 45.4, 35.8,32.0, 31.7, 30.8, 18.5, 17.4, 15.0 (×2); HRMS (ESI-TOF) m/z 511.2521([M+Na⁺], C₂₈H₄₀O₇Na requires 511.2666).

Illustrative Compound 2-140:

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 6.72 (s, 1H), 6.57 (s, 1H), 5.89-5.71(m, 2H), 5.20-5.16 (m, 4H), 5.12-4.90 (m, 4H), 4.33 (t, J=6.8 Hz, 2H),3.69 (2×q, J=7.0 Hz, 4H), 3.57 (s, 3H), 3.13 (s, 3H), 2.69-2.64 (m, 1H),2.53-2.45 (m, 4H), 2.32 (m, 2H), 2.08-2.03 (m, 2H), 1.19 (t, J=6.8 Hz,6H), 1.01 (t, J=6.5 Hz, 2H); HRMS (ESI-TOF) m/z 508.2873 ([M+H⁺],C₂₇H₄₂O₈N requires 508.2905).

Using the procedure above, the following non-limiting examples ofcompounds 2-118/2-119 shown below were prepared.

The following non-limiting examples of compounds 2-140 were prepareaccording to the procedure described here.

Example 3 General Procedure for the Metathesis Reaction

As depicted in Scheme 16, a solution of crude 2-118 or 2-119 (or mixture2-118/2-119 and 2-140 when X=Cl), in anhydrous toluene (2 mM) wastreated with Grubbs' second generation catalyst (0.10 equiv) and heatedat 80° C. for 12 h. The reaction was cooled down to room temperature andthe mixture was filtered through a pad of SiO₂, washed with CH₂Cl₂followed by a mixture EtOAc/cyclohexane 1/1 and concentrated underreduced pressure. Purification by flash chromatography (SiO₂, 0-25%EtOAc/cyclohexane gradient) afforded compounds 2-112 or 2-120 or 2-140(60-85% over two steps). Illustrative examples of compounds 2-112 or2-120 and 2-140 follow; their characterization follows respectively inthe text below each figure.

Illustrative Compounds 2-112/2-120:

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.14 (s, 1H), 6.72-6.66 (m, 1H), 5.88(d, J=15.2 Hz, 1H), 5.33-5.17 (m, 6H), 4.92-4.88 (m, 1H), 4.21 (d,J=17.0 Hz, 1H), 3.92 (d, J=17.0 Hz, 1H), 3.79-3.67 (m, 4H), 2.33-2.17(m, 5H), 2.07-1.96 (m, 2H), 1.23 (t, J=7.0 Hz, 3H), 1.21 (t, J=7.0 Hz,3H), 1.00 (d, J=5.8 Hz, 6H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 195.7,167.1, 154.7, 154.4, 147.4, 133.7, 131.2, 128.8, 128.4, 119.7, 118.0,102.7, 93.9, 93.5, 80.0, 64.8, 64.5, 44.1, 32.3, 31.2, 30.7, 30.6, 18.3,17.2, 15.0, 14.9; HRMS (ESI) m/z 517.1844 ([M+Na⁺], C₂₆H₃₅O₇ClNarequires 517.1964); [α]²⁵ _(D) +21.3 (c 1.00, CHCl₃).

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.49-7.47 (m, 2H), 7.40-7.29 (m, 3H),7.10 (s, 1H), 6.84-6.77 (m, 1H), 5.98 (d, J=15.2 Hz, 1H), 5.78 (d, J=8.8Hz, 1H), 5.44-5.30 (m, 4H), 5.15 (d, J=7.0 Hz, 1H), 5.05 (d, J=6.8 Hz,1H), 4.07 (d, J=17.0 Hz, 1H), 3.90 (d, J=17.0 Hz, 1H), 3.80 (d, J=7.0Hz, 2H), 3.60-3.51 (m, 2H), 2.68-2.62 (m, 1H), 2.50-2.47 (m, 1H),2.38-2.29 (m, 2H), 2.14-2.02 (m, 2H), 1.25 (t, J=7.0 Hz, 3H), 1.17 (t,J=7.0 Hz, 3H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 195.7, 166.7, 154.8,154.2, 147.3, 140.7, 133.3, 132.1, 128.5, 128.3 (×2), 128.2, 127.9,127.7, 126.7 (×2), 120.1, 118.1, 102.9, 93.9, 93.4, 77.4, 64.8, 64.4,44.5, 40.5, 30.7, 15.0, 14.9; HRMS (ESI) m/z 551.1807 ([M+Na⁺],C₂₉H₃₃O₇ClNa requires 551.1680); [α]²⁵ _(D) −40.4 (c 0.79, CHCl₃).

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.39-7.33 (m, 4H), 7.31-7.27 (m, 1H),6.82 (s, 1H), 6.82-6.75 (m, 1H), 6.63 (s, 1H), 6.02 (d, J=16.4 Hz, 1H),5.35-5.29 (m, 2H), 5.27-5.20 (m, 5H), 4.16 (d, J=14.6 Hz, 1H), 3.79-3.70(m, 4H), 3.52 (d, J=14.6 Hz, 1H), 3.37 (dd, J=13.4, 4.1 Hz, 1H), 2.78(dd, J=13.5, 9.4 Hz, 1H), 2.37-2.12 (m, 5H), 2.06-2.02 (m, 1H), 1.26 (t,J=7.0 Hz, 3H), 1.24 (t, J=7.0 Hz, 3H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.)δ 197.6, 167.8, 159.2, 156.5, 149.0, 137.3, 135.5, 131.8, 129.9, 129.5(×2), 128.6 (×2), 128.4, 126.7, 118.1, 109.9, 102.3, 93.5, 93.1, 75.8,64.6, 64.4, 44.4, 41.0, 36.2, 31.0, 30.6, 15.0 (×2); HRMS (ESI) m/z531.2350 ([M+Na⁺], C₃₀H₃₆O₇Na requires 531.2359); [α]²⁵ _(D) −24.1 (c0.33, CHCl₃).

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.56-7.54 (m, 2H), 7.41-7.29 (m, 3H),6.89-6.82 (m, 1H), 6.78 (d, J=2.3 Hz, 1H), 6.61 (d, J=1.8 Hz, 1H), 6.06(d, J=16.4 Hz, 1H), 5.98 (dd, J=11.7, 2.4 Hz, 1H), 5.53-5.51 (m, 2H),5.20 (d, J=7.0 Hz, 1H), 5.17 (d, J=6.4 Hz, 1H), 5.07 (d, J=7.0 Hz, 1H),4.96 (d, J=7.0 Hz, 1H), 4.20 (d, J=14.6 Hz, 1H), 3.73-3.68 (m, 2H),3.54-3.45 (m, 3H), 2.71-2.66 (m, 1H), 2.55-2.51 (m, 1H), 2.38-2.32 (m,2H), 2.23-2.06 (m, 2H), 1.22 (t, J=7.0 Hz, 3H), 1.14 (t, J=7.0 Hz, 3H);¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 197.6, 167.4, 159.3, 156.6, 149.0,140.8, 135.6, 132.2, 129.9, 128.5, 128.2 (×2), 127.9, 126.9 (×2), 117.9,109.9, 102.3, 93.2, 93.0, 76.6, 64.4, 64.3, 44.4, 40.5, 31.0, 30.6,15.0, 14.9; HRMS (ESI) m/z 517.2062 ([M+Na], C₂₉H₃₄O₇Na requires517.2197). [α]²⁵ _(D) −108.3 (c 1.00, CHCl₃).

Illustrative Compounds 2-121:

Mixture of 4 diastereoisomers: ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 6.77(s, 1H), 6.52 (s, 0.5H), 6.46 (s, 0.5H), 5.59-5.37 (m, 2H), 5.21-5.18(m, 4H), 5.09-4.92 (m, 1H), 3.75-3.70 (m, 4H), 3.53-3.48 (m, 3H),3.38-3.34 (m, 1H), 3.19-3.10 (m, 3H), 2.65-2.47 (m, 3H), 2.29-2.04 (m,6H), 1.89-1.72 (m, 2H), 1.31-1.20 (m, 6H), 1.06-0.96 (m, 6H); HRMS(ESI-TOF) m/z 544.2907 ([M+Na⁺], C₂₈H₄₃O₈NNa requires 544.2881).

Mixture of 4 diastereoisomers: ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ7.51-7.42 (m, 2H), 7.38-7.31 (m, 3H), 6.73-6.70 (m, 1H), 6.60-6.49 (m,1H), 6.45-6.31 (m, 1H), 5.73-5.39 (m, 2H), 5.23-5.00 (m, 4H), 3.75-3.69(m, 2H), 3.56-3.34 (m, 6H), 3.19-3.09 (m, 3H), 2.66-2.08 (m, 8H),1.31-1.19 (m, 5H), 1.10-1.04 (m, 3H); HRMS (ESI-TOF) m/z 578.2715([M+Na⁺], C₃₁H₄₁O₈NNa requires 578.2724).

Example 4 General Procedure for the EOM Deprotection to GenerateCompounds Deprotected-2-121 and 2-85

As depicted in Scheme 17 for compounds 2-103/2-85, to a solution of thecorresponding compound 2-120/2-112 or 2-121 (1.0 equiv) in MeOH (0.03 M)was added PS-TsOH (10.0 equiv, 3.2 mmol/g) and the suspension was shakenat 40° C. for 1 to 4 h. The reaction mixture was filtered and themethanolic solution concentrated under reduced pressure. Purification byflash chromatography (SiO₂, 0-20% EtOAc/cyclohexane gradient) affordedthe corresponding compound deprotected-2-121 or compound 2-103/2-85.(>90%). Illustrative examples of compounds deprotected-2-121 and of2-103/2-85 follow; their characterization follows respectively in thetext below each figure.

Illustrative Compound Deprotected-2-121:

Mixture of 4 diastereoisomers: ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 11.54(s, 1H), 6.33 (d, J=2.3 Hz, 1H), 6.25 (s, 1H), 5.53-5.51 (m, 1H),5.44-5.41 (m, 1H), 5.11-5.08 (m, 1H), 4.01 (d, J=11.7 Hz, 2H), 3.45 (s,3H), 3.11 (s, 3H), 2.83-2.73 (m, 1H), 2.68-2.59 (m, 1H), 2.27-2.20 (m,1H), 2.10-1.87 (m, 6H), 1.82-1.72 (m, 1H), 1.01-0.94 (m, 6H); HRMS(ESI-TOF) m/z 428.2109 ([M+Na⁺], C₂₂H₃₁O₆NNa requires 428.2044).

Illustrative Compounds 2-103/2-85:

¹H NMR (C₆D₆, 400 MHz, 25° C.) δ 12.31 (s, 1H), 6.83 (s, 1H), 6.74-6.67(m, 1H), 5.84 (bs, 1H), 5.82 (d, J=15.8 Hz, 1H), 5.03-4.95 (m, 1H),4.88-4.86 (m, 1H), 4.76-4.70 (m, 1H), 4.40 (d, J=17.6 Hz, 1H), 4.15 (d,J=17.5 Hz, 1H), 2.40-2.34 (m, 1H), 2.22-2.18 (m, 1H), 1.87-1.65 (m, 4H),1.53-1.48 (m, 1H), 0.92 (d, J=6.4 Hz, 3H), 0.66 (d, J=7.0 Hz, 3H); ¹³CNMR (C₆D₆, 100 MHz, 25° C.) δ 193.7, 164.2, 156.8, 145.8, 137.2, 131.8,129.3, 126.3, 115.3, 107.9, 103.6, 82.1, 46.4, 33.3, 30.9, 30.7, 28.8,20.1, 18.5, 18.3; HRMS (ESI-TOF) m/z 401.1170 ([M+Na⁺], C₂₀H₂₃ClO₅Narequires 401.1126); [α]²⁵ _(D) −35.6 (c 0.52, CHCl₃).

¹H NMR (C₆D₆, 400 MHz, 25° C.) δ 12.0 (bs, 1H), 7.32-7.29 (m, 3H),7.19-7.15 (m, 2H), 6.86-6.79 (m, 1H), 6.51 (d, J=2.4 Hz, 1H), 6.27-6.25(m, 1H), 6.11 (d, J=2.4 Hz, 1H), 6.02 (d, J=15.8 Hz, 1H), 5.49 (s, 1H),5.17-5.10 (m, 1H), 4.97-4.90 (m, 1H), 4.40 (d, J=16.4 Hz, 1H), 3.97 (d,J=17.2 Hz, 1H), 2.83-2.76 (m, 1H), 2.45-2.38 (m, 1H), 1.89-1.78 (m, 2H),1.67-1.58 (m, 2H); ¹³C NMR (C₆D₆, 100 MHz, 25° C.) δ 196.5, 169.6,166.1, 161.3, 146.0, 140.5, 138.8, 132.1, 130.0, 128.6 (×2), 127.3,126.6 (×2), 126.3, 112.2, 105.9, 103.0, 77.1, 48.6, 38.4, 30.9, 30.3;HRMS (ESI) m/z 401.1271 ([M+Na⁺], C₂₃H₂₂O₅Na requires 401.1359); [α]²⁵_(D) −10.3 (c 0.25, CHCl₃).

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 12.43 (s, 1H), 6.74 (d, J=1.7 Hz, 1H),6.73-6.65 (m, 1H), 6.48 (d, J=1.7 Hz, 1H), 5.92 (d, J=15.8 Hz, 1H),5.12-5.00 (m, 2H), 4.91-4.80 (m, 1H), 4.19 (d, J=17.0 Hz, 1H), 3.84 (d,J=16.4 Hz, 1H), 2.77 (m, 1H), 2.64-2.57 (m, 1H), 2.01-1.97 (m, 1H),1.89-1.70 (m, 3H), 1.61-1.56 (m, 2H), 1.30-1.21 (m, 2H), 0.90 (t, J=6.7Hz, 3H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 197.5, 169.9, 165.6, 160.6,147.5, 140.2, 131.9, 129.5, 127.0, 112.8, 106.1, 102.9, 76.2, 48.7,35.7, 34.3, 31.1, 29.7, 19.4, 13.8; HRMS (ESI-TOF) m/z 367.1330([M+Na⁺], C₂₀H₂₄O₅Na requires 367.1521); [α]²⁵ _(D) +21.6 (c 0.36,CHCl₃).

¹H NMR (CD₃OD, 400 MHz, 25° C.) δ 6.78-6.71 (m, 1H), 6.29 (d, J=2.4 Hz,1H), 6.22 (d, J=2.0 Hz, 1H), 5.87 (d, J=15.5 Hz, 1H), 5.37-5.23 (m, 3H),4.01 (d, J=17.2 Hz, 1H), 3.92 (d, J=17.0 Hz, 1H), 2.67-2.61 (m, 1H),2.29-2.15 (m, 5H), 1.31 (d, J=6.4 Hz, 3H); ¹³C NMR (CD₃OD, 100 MHz, 25°C.) δ 198.5, 169.8, 164.2, 162.3, 148.4, 139.1, 131.6, 129.6, 127.3,111.7, 101.7, 72.0, 47.7, 36.8, 30.8, 30.7, 17.4, (1 quartenary carbonis not visible); HRMS (ESI) m/z 339.1141 ([M+Na], C₁₈H₂₀O₅Na requires339.1203). [α]²⁵ _(D) −45.1 (c 0.27, CHCl₃).

¹H NMR (CD₃OD, 400 MHz, 25° C.) δ 6.74-6.68 (m, 1H), 6.48 (s, 1H), 5.86(d, J=15.2 Hz, 1H), 5.31-5.25 (m, 2H), 4.39 (t, J=5.3 Hz, 2H), 4.27 (s,2H), 2.43-2.40 (m, 2H), 2.25 (m, 4H); ¹³C NMR (CD₃OD, 100 MHz, 25° C.) δ196.9, 170.1, 161.9, 158.1, 147.8, 135.9, 130.9, 130.2, 129.9, 115.2,107.3, 102.4, 65.9, 46.2, 31.3, 30.9, 30.5; HRMS (ESI) m/z 337.0797([M+H⁺], C₁₇H₁₈O₅Cl requires 337.0837).

Example 5 General Procedure for the Synthesis of Compounds 2-141

As depicted in Scheme 17, to a solution of corresponding compound2-120/2-112 (1.0 equiv) in MeOH (0.03 M) at 0° C. was added BER-resin(Borohydride on Amberlite®, 1.0 equiv, 2.5 mmol g⁻¹) and the reactionwas stirred over 12 h. The reaction was then filtered and concentratedunder reduced pressure. Purification by flash chromatography (SiO₂,0-20% EtOAc/cyclohexane gradient) afforded 2-141 (˜60%) as a mixture oftwo diastereoisomers (1:1). An illustrative example of compound 2-141follows; its characterization data are in the text below the figure.

Illustrative Compound 2-141:

¹H NMR (CD₃Cl, 400 MHz) δ 7.05 (s, 1H), 6.99 (s, 1H), 5.64-5.57 (m, 2H),5.54-5.53 (m, 2H), 5.49-5.35 (m, 6H), 5.31-5.28 (m, 4H), 5.24-5.16 (m,4H), 5.13-5.08 (m, 1H, 35′), 4.68 (m, 1H, 35′), 4.56 (m, 1H, 35),3.81-3.69 (m, 8H), 3.25 (dd, J=13.9, 8.0 Hz, 1H, 35), 3.19 (dd, J=13.7,4.8 Hz, 1H, 35′), 3.11 (dd, J=13.5, 10.1 Hz, 1H, 35′), 2.90 (dd, J=13.9,5.12 Hz, 1H, 35), 2.35 (m, 9H), 2.09-1.95 (m, 1H), 1.80-1.70 (m, 2H),1.39 (d, J=2.9 Hz, 3H, 35), 1.37 (d, J=3.2 Hz, 3H, 35′), 1.24 (2×q,J=6.9 and 5.0 Hz, 12H, 35+35′); HRMS (ESI) m/z 491.1729 ([M+Na⁺],C₂₄H₃₃ClO₇Na requires 491.1807).

Example 6 General Procedure for the Synthesis of Compounds 2-142

As depicted in Scheme 17, to a solution of the corresponding compound2-141 (1.0 equiv) in MeOH (0.02 M) was added PS-TsOH (10.0 equiv, 3.2mmol g⁻¹) and the suspension was shaken at 40° C. for 4 h. The reactionmixture was then filtered and the methanolic solution concentrated underreduced pressure. Purification by preparative TLC (SiO₂, 25%EtOAc/cyclohexane) afforded 2-142 (˜90%) as a mixture of twodiastereoisomers (1:1). An illustrative example of compound 2-142follows; its characterization data are in the text below the figure.

Illustrative Compound 2-142:

¹H NMR ((CD₃)₂CO, 400 MHz) δ 12.30 (s, 2H), 11.43 (s, 2H), 6.75 (s, 2H),6.00 (bdd, J=6.4, 6.2 Hz, 1H), 5.97 (bdd, J=6.4, 6.2 Hz, 1H), 5.97 (bd,J=6.7 Hz, 1H), 5.77 (bd, J=6.7 Hz, 1H), 5.57-5.48 (m, 4H), 5.18-5.14 (m,2H), 3.38-3.28 (m, 3H), 3.02 (dd, J=16.1, 10.5 Hz, 1H), 2.41-2.09 (m,12H), 1.11 (d, J=6.2 Hz, 6H); HRMS (ESI) m/z 375.1029 ([M+Na⁺],C₁₈H₂₁ClO₅Na requires 375.0970).

Example 7 General Procedure for the Synthesis of Compounds 2-143

As depicted in Scheme 17, to a solution of the corresponding compound2-141 (1.0 equiv) in DMF (0.02 M) were added Ac₂O (1.2 equiv),morpholinomethyl polystyrene (1.2 equiv, 3.2 mmol g⁻¹) and DMAP (0.05equiv) at 23° C. and the mixture was stirred for 30 min, followed by TLCuntil consumption of the starting material. Then, the resin was filteredand the organic phase was concentrated under reduced pressure.Purification by PTLC (SiO₂, 20% EtOAc/cyclohexane) affordedcorresponding 2-143 (˜80%) as a mixture of two diastereoisomers 1:1: Anillustrative example of compound 2-143 follows; its characterizationdata are in the text below the figure.

Illustrative Compound 2-143:

¹H NMR (CD₃Cl, 400 MHz) δ 7.04 (s, 1H), 7.01 (s, 1H), 5.86 (dd, J=15.0,6.9 Hz, 1H), 5.67 (dd, J=12.4, 6.2 Hz, 1H), 5.60-5.54 (m, 4H), 5.48 (dd,J=7.2, 7.2 Hz, 1H), 5.41-5.34 (m, 3H), 5.32-5.30 (m, 4H), 5.28-5.23 (m,2H), 5.21 (dd, J=11.0, 6.7 Hz, 2H), 5.17 (dd, J=11.8, 6.9 Hz, 2H),3.81-3.69 (m, 8H), 3.43 (dd, J=14.2, 7.5 Hz, 1H), 3.23-3.15 (m, 2H),2.85 (dd, J=13.9, 5.4 Hz, 1H), 2.30-2.17 (m, 8H), 2.12 (s, 3H), 2.06 (s,3H), 1.95-2.00 (m, 4H), 1.39 (2×d, J=5.6 Hz, 6H), 1.24 (m, 12H); HRMS(ESI) m/z 533.1864 ([M+Na⁺], C₂₆H₃₅ClO₈Na requires 533.1913).

Example 8 General Procedure for the Synthesis of Compounds 2-144

As depicted in Scheme 17, to a solution of corresponding compound 2-143(1.0 equiv) in MeOH (0.02 M) was added PS-TsOH (10.0 equiv, 3.2 mmol/g)and the suspension was shaken at 40° C. for 4 h. The reaction mixturewas filtered and the methanolic solution concentrated under reducedpressure. Purification by PTLC (SiO₂, 20% EtOAc/cyclohexane) affordedcompounds 2-144 (˜60% yield). An illustrative example of compound 2-144follows; its characterization data are in the text below the figure.

Illustrative Compound 2-144:

Mixture of diastereoisomers 2:1: ¹H NMR (CD₃Cl, 400 MHz) δ 12.6 (s, 1H),12.12 (s, 0.5H), 6.93 (d, J=8.7 Hz, 0.5H), 6.66 (s, 1H), 6.64 (s, 0.5H),6.62-6.60 (m, 1H), 6.10-6.05 (m, 3H), 5.47-5.33 (m, 4.5H), 2.60-2.53 (m,1.5H), 2.26-2.02 (m, 7.5H), 1.44 (d, J=6.2 Hz, 1.5H), 1.43 (d, J=6.4 Hz,3H); HRMS (ESI) m/z 357.0898 ([M+Na⁺], C₁₈H₁₉ClO₄Na requires 357.0864).

Example 9 General Procedure for the Synthesis of Compounds 2-145

As depicted in Scheme 18, to a solution of corresponding compound 2-85(1.0 equiv) in methanol (0.03 M) was added sulfamic acid resin (10.0equiv) and the suspension was stirred for 15 h at 40° C. with. Thereaction was then filtered, the resin washed several times with CH₂Cl₂.Concentration under reduced pressure followed by purification on PTLC(Hexane/EtOAc: 1/1) afforded desired compounds 2-145 as a mixturediastereoisomers (2:1). An illustrative example of compound 2-145follows; its characterization data are in the text below the figure.

Illustrative Compound 2-145:

¹H NMR (C₆D₆, 400 MHz, 25° C.) δ 12.28 (s, 0.4H), 11.91 (s, 0.6H),7.21-7.11 (m, 5H), 6.62 (s, 1H), 6.03-6.01 (m, 1H), 5.58 (bs, 1H),5.38-5.33 (m, 1H), 5.27-5.20 (m, 1H), 4.76 (d, J=17.5 Hz, 0.6H), 4.02(d, J=17.0 Hz, 0.4H), 4.18 (d, J=18.1 Hz, 0.6H), 4.09 (d, J=17.0 Hz,0.4H), 3.87 (bs, 0.4H), 3.81 (bs, 0.6H), 3.15 (s, 1.8H), 3.12 (s, 1.2H),2.83-2.78 (m, 1H), 2.45-2.30 (m, 2H), 2.18-2.16 (m, 1H), 2.02-1.97 (m,2H), 1.79-1.72 (m, 2H); HRMS (ESI-TOF) m/z 467.1366 ([M+Na⁺],C₂₄H₂₅O₆ClNa requires 467.1232).

Example 10 General Procedure for the Synthesis of Compounds 2-146

As depicted in Scheme 19, to a solution of corresponding compound2-103/2-85 (1.0 equiv) in CH₂Cl₂/AcOH 10/1 (0.08 M)(polystyrylmethyl)trimethylammonium cyanoborohydride (2.0 equiv, 3.5mmol g⁻¹) was added at 23° C. and the reaction was monitored by TLCuntil the starting material had been consumed (4 h). Then, the resin wasfiltered and the organic phase was concentrated under reduced pressure.Purification by PTLC (SiO₂, 30% EtOAc/cyclohexane) afforded compounds2-146 (50-60%). An illustrative example of compound 2-146 follows; itscharacterization data are in the text below the figure.

Illustrative Compound 15:

¹H NMR (CD₃Cl, 400 MHz) δ 11.75 (s, 1H), 6.65 (s, 1H), 5.48 (m, 2H),5.49 (ddt, J=6.1, 3.5, 2.9 Hz, 1H), 4.53 (d, J=17.5 Hz, 1H), 4.04 (d,J=17.7 Hz, 1H), 2.61-2.54 (m, 2H), 2.48-2.28 (m, 3H), 2.19-2.14 (m, 1H),2.08-1.99 (m, 1H), 1.72-1.61 (m, 3H), 1.41 (d, J=6.4 Hz, 3H); HRMS (ESI)m/z 375.1050 ([M+Na⁺], C₁₈H₂₁ClO₅Na requires 375.0970).

Example 11 General Procedure for the Synthesis of Compounds 2-147

As depicted in Scheme 19, to a solution of corresponding compound2-103/2-85 (1.0 equiv), in THF (0.05 M) were added in a sequentialmanner the corresponding alcohol (2.0 equiv), triphenylphosphine (2.0equiv) and ethoxycarbonylazocarboxymethyl polystyrene (2.0 equiv, 1.3mmol g⁻¹). The reaction mixture was shaken at room temperature for 8hours, and then, the resin was filtered and the filtrates were directlypurified by PTLC (SiO₂, 10% EtOAc/cyclohexane) to afford a mixture ofcompound 2-147 along with the bis-allylated product (78%). Anillustrative example of compound 2-147 follows; its characterizationdata are in the text below the figure.

Illustrative Compound 2-147:

Mixture with the corresponding bis-allylated compound (1:1): ¹H NMR(CD₃Cl, 400 MHz) δ 11.83 (s, 1H), 6.82 (ddd, J=15.7, 8.2, 4.6 Hz, 1H),6.72-6.65 (m, 1H), 6.46 (s, 1H), 6.41 (s, 1H), 6.09-5.98 (m, 3H), 5.82(d, J=15.7 Hz, 1H), 5.46-5.16 (m, 8H), 4.57-4.54 (m, 3H), 4.51-4.49 (m,3H), 4.19 (d, J=17.5 Hz, 1H), 4.11 (d, J=14.6 Hz, 1H), 3.78 (d, J=17.0Hz, 1H), 3.51 (d, J=14.2 Hz, 1H), 2.76-2.69 (m, 1H), 2.38-2.05 (m, 11H),1.42 (d, J=6.2 Hz, 3H), 1.35 (d, J=6.3 Hz, 3H); mono-allylated compoundHRMS (ESI) m/z 413.1103 ([M+Na⁺], C₂₁H₂₃ClO₅Na requires 413.1132);bis-allylated compound HRMS (ESI) m/z 453.1422 ([M+Na⁺], C₂₄H₂₇ClO₅Narequires 453.1449).

Example 12 General Procedure for the Synthesis of Compounds 2-148

As depicted in Scheme 19, to a solution of the corresponding compound2-103/2-85 (1.0 equiv) in CH₂Cl₂ (0.05 M) was added TBD-methylpolystyrene (2.0 equiv, 2.9 mmol g⁻¹) and the corresponding alkylbromide or chloride (BrCH₂COO^(t)Bu, EOMCl) (0.9 equiv) at 23° C. andthe mixture was shaken for 3 h. The resin was then filtered and thefiltrates were concentrated under reduced pressure. Purification by PTLC(SiO₂, 30% EtOAc/cyclohexane) afforded corresponding compound 2-148(>90%). Illustrative examples of compound 2-148 follows; theircharacterization data are presented respectively in the text below eachfigure.

Illustrative Compounds 2-148:

¹H NMR (CD₃Cl, 400 MHz) δ 11.84 (s, 1H), 6.69 (m, 1H), 6.41 (s, 1H),5.76 (d, J=15.0 Hz, 1H), 5.43 (m, 1H), 5.26 (ddd, J=15.0, 9.1, 4.8 Hz,1H), 5.18-5.11 (m, 1H), 4.65 (s, 2H), 4.33 (d, J=17.7 Hz, 1H), 4.16 (d,J=17.5 Hz, 1H), 2.65-2.58 (m, 1H), 2.37-2.34 (m, 2H), 2.25-2.21 (m, 1H),2.12-2.01 (m, 2H), 1.53 (s, 9H), 1.34 (d, J=6.5 Hz, 3H); HRMS (ESI) m/z487.1498 ([M+Na⁺], C₂₄H₂₉ClO₇Na requires 487.1494).

¹H NMR (C₆D₆, 400 MHz, 25° C.) δ 11.76 (s, 1H), 6.86 (s, 1H), 6.70 (dt,J=14.9, 7.3 Hz, 1H), 5.77 (d, J=15.8 Hz, 1H), 5.46-5.42 (m, 1H), 5.37(s, 2H), 5.30-5.19 (m, 2H), 4.34 (d, J=17.6 Hz, 1H), 4.16 (d, J=18.1 Hz,1H), 3.80 (q, J=7.0 Hz, 2H), 2.66-2.59 (m, 1H), 2.37-2.34 (m, 2H),2.26-2.21 (m, 1H), 2.13-2.06 (m, 2H), 1.34 (d, J=6.4 Hz, 3H), 1.27 (t,J=7.0 Hz, 3H); HRMS (ESI) m/z ([M+Na], C₂₁H₂₅O₆ClNa requires 431.1237).

Example 13 General Procedure for the Synthesis of Compounds 2-149

As depicted in Scheme 19, to a solution of compound 2-103/2-85 (1.0equiv) in acetone/H₂O 10/1 (0.05 M) was added OsO₄ (0.1 equiv) followedby NMO (1.0 equiv) at 23° C. and the mixture was stirred for 1 h. Thecrude mixture was filtered through a plug of silica, concentrated andpurified by PTLC (SiO₂, 30% EtOAc/cyclohexane) to afford 2-149 (>70%) asa mixture of two diastereoisomers. An illustrative example of compound2-149 follows; its characterization data are in the text below thefigure.

Illustrative Compound 2-149:

¹H NMR (CD₃OD, 400 MHz) δ 7.19 (m, 1H), 6.89-6.81 (m, 1H), 6.52 (s, 1H),6.47 (s, 1H), 6.20 (d, J=16.1 Hz, 1H), 6.04 (d, J=15.6 Hz, 1H),5.54-5.49 (m, 1H), 5.43-5.36 (m, 1H), 4.50 (d, J=17.7 Hz, 1H), 4.46 (d,J=17.7 Hz, 1H), 4.39 (d, J=17.2 Hz, 1H), 4.07 (d, J=17.2 Hz, 1H),3.80-3.64 (m, 2H), 3.51-3.46 (m, 2H), 2.62-2.58 (m, 2H), 2.39-2.30 (m,2H), 2.27-2.18 (m, 2H), 2.08-2.98 (m, 2H), 2.00-1.85 (m, 4H), 1.44 (d,J=6.4 Hz, 6H); HRMS (ESI) m/z 407.1031 ([M+Na⁺], C₁₈H₂₁ClO₇Na requires407.0868).

Example 14 General Procedure for the Synthesis of Compounds 2-150

As depicted in Scheme 19, to a solution of compound 2-103/2-85 (1.0equiv) in CH₃CN (0.03 M) at 0° C. was added freshly made DMDO (1.2equiv, 0.04 M in acetone) and the mixture was stirred for 30 min. Afterevaporation of the solvents under reduced pressure, purification by PTLC(SiO₂, 30% EtOAc/cyclohexane) afforded epoxides 2-150 (>90%) as amixture of two diastereoisomers. Illustrative examples of compound 2-150follow; their characterization data are presented respectively in thetext below each figure.

Illustrative Compounds 2-150:

¹H NMR (CDCl₃, 400 MHz) δ 11.84 (s, 2H), 6.94-6.82 (m, 2H), 6.69 (s,1H), 6.66 (s, 1H), 6.23 (d, J=17.1 Hz, 1H), 6.11 (dd, J=13.2, 1.6 Hz,1H), 5.39 (tdd, J=7.5, 3.2, 2.7 Hz, 1H), 5.32 (m, 1H), 4.53 (d, J=17.7Hz, 2H), 4.27 (d, J=17.7 Hz, 2H), 2.79-2.76 (m, 1H), 2.74-2.69 (m, 1H),2.58 (m, 1H), 2.56 (m, 1H), 2.47-2.24 (m, 8H), 2.13-2.08 (m, 1H),2.05-2.03 (m, 1H), 1.91 (dd, J=4.3, 4.3 Hz, 1H), 1.87 (dd, J=4.3, 4.3Hz, 1H), 1.51 (d, J=6.4 Hz, 3H), 1.35 (d, J=6.4 Hz, 3H); HRMS (ESI) m/z389.0724 ([M+Na⁺], C₁₈H₁₉ClO₆Na requires 389.0762).

¹H NMR (C₆D₆, 400 MHz, 25° C.) δ 11.56 (2×s, 2H), 6.92-6.82 (m, 2H),6.71 (s, 1H), 6.67 (s, 1H), 6.20 (m, 3H), 6.06 (d, J=15.8 Hz, 1H), 5.11(bs, 1H), 5.94 (m, 1H), 4.46 (2×d, J=18.1 Hz, 2H), 4.20 (2×d, J=18.1 Hz,2H), 2.72-2.70 (m, 2H), 2.53-2.48 (m, 4H), 2.38-2.35 (m, 3H), 2.25-2.13(m, 5H), 1.84-1.77 (m, 2H), 1.05-1.01 (m, 6H), 0.91-0.88 (m, 3H),0.86-0.84 (m, 3H); HRMS (ESI) m/z 417.1128 ([M+Na⁺], C₂₀H₂₃O₆ClNarequires 417.1075).

¹H NMR (C₆D₆, 400 MHz, 25° C.) δ 11.80 (2×s, 2H), 7.43-7.18 (m, 10H),7.03-6.95 (m, 2H), 6.69 (s, 1H), 6.61 (s, 1H), 6.30 (d, J=16.4 Hz, 1H),6.21 (d, J=15.8 Hz, 1H), 6.15-6.10 (m, 1H), 6.03 (d, J=11.1 Hz, 1H),4.84 (2×d, J=18.1 Hz, 2H), 4.41 (2×d, J=17.6 Hz, 2H), 2.68-2.60 (m, 4H),2.41-2.27 (m, 8H), 1.83-1.76 (m, 4H); HRMS (ESI) m/z 451.1028 ([M+Na],C₂₃H_(21l O) ₆ClNa requires 451.0919).

Major isomer: ¹H NMR (C₆D₆, 400 MHz, 25° C.) δ 11.94 (s, 1H), 7.36-7.28(m, 5H), 6.95-6.88 (m, 1H), 6.42 (s, 1H), 6.22 (s, 1H), 6.11 (d, J=15.8Hz, 1H), 5.47 (m, 1H), 5.41 (bs, 1H), 4.43 (d, J=17.5 Hz, 1H), 3.56 (d,J=17.6 Hz, 1H), 3.19 (dd, J=13.7, 6.0 Hz, 1H), 3.03 (dd, J=13.7, 7.9 Hz,1H), 2.87 (bs, 1H), 2.70-2.28 (m, 4H), 2.03-1.93 (m, 2H); HRMS (ESI) m/z431.1578 ([M+Na], C₂₄H₂₄O₆Na requires 431.1465).

¹H NMR (C₆D₆, 400 MHz, 25° C.) δ 11.98 (s, 1H), 6.91-6.83 (m, 1H), 6.43(d, J=2.3 Hz, 1H), 6.24 (d, J=2.4 Hz, 1H), 6.11 (d, J=15.8 Hz, 1H), 5.35(bs, 1H), 5.29 (m, 1H), 4.52 (d, J=17.5 Hz, 1H), 3.63 (d, J=17.5 Hz,1H), 2.77 (m, 2H), 2.57-2.52 (m, 2H), 2.46-2.27 (m, 2H), 2.14-2.10 (m,1H), 1.93-1.88 (m, 1H), 1.48 (d, J=6.4 Hz, 3H); other isomer: ¹H NMR(C₆D₆, 400 MHz, 25° C.) δ 11.67 (s, 1H), 6.89-6.83 (m, 1H), 6.40 (d,J=2.4 Hz, 1H), 6.24 (d, J=2.9 Hz, 1H), 6.21 (d, J=16.4 Hz, 1H), 5.37(bs, 1H), 5.22 (m, 1H), 4.20 (d, J=17.0 Hz, 1H), 4.06 (d, J=17.0 Hz,1H), 2.74 (m, 2H), 2.57-2.20 (m, 4H), 1.80-1.76 (m, 1H), 1.68-1.60 (m,1H), 1.37 (d, J=6.4 Hz, 3H); HRMS (ESI) m/z 355.1249 ([M+Na], C₁₈H₂₀O₆Narequires 355.1152).

Example 15 General Procedure for the Synthesis of Compounds 2-151

As depicted in Scheme 20, to a solution of compound 2-120 (1.0 equiv) indioxane (0.05 M) at 23° C. was added HCl_(conc.) (20 equiv), and themixture was stirred for 3 h. After that time the reaction was filteredthrough a plug of silica gel, the solvents were evaporated under reducedpressure, and purified by PTLC (SiO₂, 30% EtOAc/cyclohexane) to affordcompound 2-151 (>75%) as a mixture of two diastereoisomers. Illustrativeexamples of compound 2-151 follow; their characterization data arepresented respectively in the text below each figure.

Illustrative Compounds 2-151:

¹H NMR (CDCl₃, 400 MHz) δ 12.11 (s, 1H), 11.78 (s, 1H), 6.51 (s, 1H),6.43 (s, 1H), 6.41 (d, J=2.4 Hz, 1H), 6.37 (d, J=2.7 Hz, 1H), 6.21 (d,J=2.4 Hz, 1H), 6.11 (d, J=2.4 Hz, 1H), 5.59-5.51 (m, 3H), 5.40-5.32 (m,3H), 4.54 (d, J=17.2 Hz, 1H), 4.42 (d, J=17.2 Hz, 1H), 3.60 (d, J=17.2Hz, 1H), 3.45 (d, J=17.0 Hz, 1H), 3.28 (dd, J=18.5, 9.4 Hz, 1H), 3.11(dd, J=13.7, 6.2 Hz, 1H), 3.07 (dd, J=13.4, 4.6 Hz, 1H), 2.76 (dd,J=19.0, 6.2 Hz, 1H), 2.62 (ddd, J=15.5, 8.8, 4.0 Hz, 1H), 2.54 (ddd,J=15.3, 6.2, 3.2 Hz, 1H), 2.40-2.26 (m, 4H), 2.25-2.13 (m, 4H),2.03-1.91 (m, 2H), 1.42 (d, J=6.4 Hz, 3H), 1.40 (d, J=6.4 Hz, 3H); HRMS(ESI) m/z 375.0928 ([M+Na⁺], C₁₈H₂₁ClO₅Na requires 375.0970).

¹H NMR (C₆D₆, 400 MHz, 25° C.) δ 11.76 (s, 0.5H), 11.36 (s, 0.5H),7.40-7.29 (m, 5H), 6.65 (s, 0.5H), 6.62 (s, 0.5H), 6.18 (t, J=5.8 Hz,1H), 6.14 (s, 0.5H), 6.12 (s, 0.5H), 5.67-5.62 (m, 1H), 5.55-5.49 (m,1H), 4.93 (d, J=18.1 Hz, 0.5H), 4.80 (d, J=17.1 Hz, 0.5H), 4.58-4.56 (m,1H), 4.38 (d, J=18.1 Hz, 0.5H), 4.18 (d, J=17.1 Hz, 0.5H), 3.33-3.27 (m,1H), 3.10 (dd, J=18.4, 3.8 Hz, 0.5H), 2.84-2.68 (m, 2.5H), 2.42-2.32 (m,2H), 2.23-2.17 (m, 1H), 2.13-2.04 (m, 1H); HRMS (ESI-TOF) m/z 471.0754([M+Na⁺], C₂₃H₂₂O₅Cl₂Na requires 471.0737).

Example 16 General Procedure for the Elimination of β-Cl from Compounds2-151

As depicted in Scheme 20, to a solution of compound 2-151 (95 mg, 270mmol) in CH₂Cl₂ (5 ml) at 23° C. PS-TBD (51 mg, 2.6 mmol/g) was added,and the mixture was stirred for 8 hours. After that time the reactionwas filtered, the solvents were evaporated under reduced pressure, andpurification by flash chromatography (SiO₂, 0-30% EtOAc/cyclohexanegradient) afforded 2-103 (X=Cl, R=Me) (84 mg, 98%).

Example 17 General Procedure for the Synthesis of Compounds 2-153

As depicted in Scheme 20, to a solution of compound 2-103 (X=Cl, R=Me)(12.9 mg, 40.8 mmol) in CH₂Cl₂ (1 ml) at 23° C. DHP (3.7 μL, 40.8 μmol)and PS-TsOH (12.7 mg, 40.8 μmol, 3.2 mmol/g) were added, and the mixturewas stirred for 5 hours. After that time the reaction was filtered andthe solvents were evaporated under reduced pressure. Purification byPTLC (SiO₂, 30% EtOAc/cyclohexane) afforded 2-153 (13.8 mg, 85%) as amixture of two diastereoisomers. An illustrative example of compounds2-153 follows; its characterization data are in the text below thefigure.

Illustrative Compound 2-153:

¹H NMR (CDCl₃, 400 MHz) δ 12.33 (s, 1H), 12.11 (s, 1H), 9.45 (s, 1H),9.40 (s, 1H), 6.67, (m, 2H), 6.28 (2×s, 2H), 5.83 (d, J=13.2 Hz, 1H),5.79 (d, J=12.9 Hz, 1H), 5.35-5.30 (m, 3H), 5.27-5.22 (m, 3H), 5.06 (bd,J=8.2 Hz, 2H), 4.10 (d, J=17.5 Hz, 2H), 3.90-3.85 (m, 1H), 3.80-3.76 (m,1H), 3.65 (d, J=17.7 Hz, 2H), 3.57-3.52 (m, 2H), 3.46-3.41 (m, 2H),2.77-2.71 (m, 3H), 2.53-2.49 (m, 3H), 2.36-2.29 (m, 4H), 2.24-1.56 (m,12H), 1.31 (d, J=6.4 Hz, 3H), 1.28 (d, J=6.4 Hz, 3H); HRMS (ESI) m/z423.1778 ([M+Na⁺], C₂₃H₂₈O₆Na requires 423.1778.

Example 18 General Procedure for the Synthesis of Compounds 2-154

As depicted in Scheme 20, to a solution of corresponding compound 2-120(1.0 equiv) in pyridine/AcOH (5/1, 0.03 M) was added the correspondinghydroxylamine (5.0 equiv) and the mixture was heated up to 40° C. Afterstirring overnight the solvents were evaporated under reduced pressurewith SiO₂. Elution of the compound over a short path of SiO₂ with amixture of 30% EtOAc/cyclohexane afforded after evaporation 2-154 (˜99%)as a mixture of two diastereoisomers cis/trans). An illustrative exampleof compound 2-154 follows; its characterization data are in the textbelow the figure.

Illustrative Compound 2-154:

¹H NMR (CDCl₃, 400 MHz) δ 7.50-7.25 (m, 10H), 6.82 (s, 1H), 6.75 (s,1H), 6.66 (s, 1H), 6.48 (s, 1H), 6.24-6.11 (m, 2H), 6.11-6.05 (m, 2H),5.45-5.38 (m, 4H), 5.34-5.31 (m, 14H), 4.50 (d, J=17.2 Hz, 1H),3.38-3.65 (m, 8H), 3.60 (d, J=17.1 Hz, 1H), 3.54 (d, J=17.1 Hz, 1H),3.24 (d, J=17.2 Hz, 1H), 2.48-2.36 (m, 4H), 2.17-2.21 (m, 2H), 2.04-2.11(m, 2H), 1.95-1.83 (m, 2H), 1.62-1.51 (m, 2H), 1.49 (d, J=6.4 Hz, 6H),1.20-1.32 (m, 12H); ¹³C NMR (CDCl₃, 100 MHz) δ 168.02, 167.85, 159.08,158.83, 157.23, 155.55, 155.36, 154.19, 140.75, 138.23, 138.19, 137.75,136.93, 136.74, 132.32, 132.28, 128.34 (×2), 128.31 (×2), 128.18, 128.09(×2), 127.99 (×2), 127.71, 127.63, 125.50, 118.82, 118.56, 118.34,108.84, 108.50, 101.72, 101.68, 93.49, 93.44, 93.12 (×2), 77.21, 76.02,75.88, 71.18, 70.99, 64.47, 64.45, 64.33, 64.31, 39.99, 39.96, 34.87,32.42, 32.31, 31.63, 31.09, 28.86, 20.25, 20.19, 15.04 (×2), 14.98 (×2);HRMS (ESI) m/z 560.2627 ([M+Na⁺], C₃₁H₃₉NO₇Na requires 560.2619.

Example 19 General Procedure for the Synthesis of Compounds 2-155

As depicted in Scheme 20, to a solution of compound 2-154 (1.0 equiv) inMeOH (0.02 M) was added PS-TsOH (10.0 equiv, 3.2 mmol/g) and thesuspension was shaken at 40° C. for 4 h. The reaction mixture wasfiltered and the methanolic solution concentrated under reducedpressure. The crude product obtained was submitted without furtherpurification to the next step. Thus to a solution in CH₂Cl₂ (0.02 M) ofthis crude at 23° C. were added DHP (1.0 equiv) and PS-TsOH (cat, 3.2mmol/g) and the mixture was stirred for 5 hours. After that time themixture was filtered, the solvents were evaporated under reducedpressure, and purification by PTLC (SiO₂, 30% EtOAc/cyclohexane)afforded two different diastereoisomers 1:1 of 2-155 (˜65%). Anillustrative example of compound 2-155 follows; its characterizationdata are in the text below the figure.

Illustrative Compound 2-155:

Less polar diastereoisomers: ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 9.25 (s,1H), 9.24 (s, 1H), 7.46-7.33 (m, 10H), 6.29 (s, 1H), 6.26 (s, 1H),6.07-6.02 (m, 2H), 5.75 (d, J=15.8 Hz, 1H), 5.69 (d, J=15.8 Hz, 1H),5.44-5.38 (m, 6H), 5.23 (s, 4H), 5.03 (d, J=8.8 Hz, 2H), 4.34-4.13 (m,6H), 3.69-3.63 (m, 2H), 2.70-2.67 (m, 2H), 2.30-2.16 (m, 6H), 2.08-1.94(m, 8H), 1.73-1.65 (m, 8H), 1.42 (t, J=6.4 Hz, 3H), 1.39 (t, J=7.0 Hz,3H); HRMS (ESI) m/z 528.2562 ([M+Na⁺], C₃₀H₃₅NO₆Na requires 528.2357.

More polar diastereoisomers: ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 11.61 (s,1H), 9.27 (s, 1H), 7.41-7.33 (m, 5H), 6.62 (d, J=16.4 Hz, 1H), 6.47 (s,1H), 6.15-6.07 (m, 1H), 5.50-5.38 (m, 3H), 5.16 (s, 2H), 5.04 (d, J=10.5Hz, 1H), 4.30 (d, J=15.2 Hz, 1H), 4.24 (d, J=10.5 Hz, 1H), 3.84 (d,J=15.2 Hz, 1H), 3.66 (t, J=11.4 Hz, 1H), 2.71-2.65 (m, 1H), 2.28-2.08(m, 6H), 1.73-1.64 (m, 5H), 1.38 (t, J=7.0 Hz, 3H); HRMS (ESI) m/z528.2494 ([M+Na⁺], C₃₀H₃₅NO₆Na requires 528.2357.

Example 20 General Procedure for the Synthesis of Compounds 2-128

As depicted in Scheme 21, to a solution of pochonin D (2-85, X=Cl andR=Me) (25 mg, 71.2 μmol) in DMF (5 ml) TBSCl (53.6 mg, 356 μmol) andimidazole (23.6 mg, 356 μmol) were added and the mixture was stirred for3 hours at room temperature. Purification by column chromatography (SiO₂0-30% EtOAc/cyclohexane gradient) afforded after evaporation 2-128 (40mg, 98%). An illustrative example of compound 2-128 follows; itscharacterization data are in the text below the figure.

Illustrative Compound 2-128:

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 6.71 (dt, J=15.3, 7.3 Hz, 1H), 6.45(s, 1H), 5.81 (d, J=15.3 Hz, 1H), 5.25 (s, 2H), 5.04-5.03 (m, 1H), 3.89(d, J=17.4 Hz, 1H), 3.57 (d, J=17.4 Hz, 1H), 2.31-2.04 (m, 6H), 1.35 (d,J=6.4 Hz, 3H), 1.03 (s, 9H), 0.99 (s, 9H), 0.28-0.24 (m, 12H); ¹³C NMR(CDCl₃, 100 MHz, 25° C.) δ 195.8, 166.8, 152.9, 151.7, 146.5, 132.7,131.9, 128.6, 126.8, 122.8, 119.7, 110.7, 71.9, 45.6, 38.5, 30.9, 25.7(×4), 25.6 (×4), 18.7, 18.3, −4.1 (×2), −4.4 (×2); HRMS (ESI) m/z601.2568 ([M+Na], C₃₀H₄₇ClO₅Si₂Na requires 601.2543).

Example 21 General Procedure for the Synthesis of Compounds 2-157

As depicted in Scheme 21, to a solution of compound 2-128 (1.0 equiv) inpyridine/AcOH (5/1, 250 μL) was added the corresponding hydroxylamine(5.0 equiv) and the mixture was heated up to 40° C. After stirringovernight the solvents were evaporated under reduced pressure, andfiltration on SiO₂ with a mixture of 30% EtOAc/cyclohexane affordedafter evaporation two isomers of 2-157 ˜90%. An illustrative example ofcompound 2-157 follows; its characterization data are in the text belowthe figure.

Illustrative Compound 2-157:

cis oxime ¹H NMR (CDCl₃, 400 MHz) δ 7.42 (bd, J=6.4 Hz, 2H), 7.36 (bdd,J=7.5, 6.9 Hz, 2H), 7.34-7.32 (m, 1H), 6.52 (d, J=16.1 Hz, 1H), 6.38 (s,1H), 6.18-6.10 (m, 1H), 5.36-5.32 (m, 2H), 5.16 (bs, 2H), 4.99-4.95 (m,1H), 3.79-3.76 (m, 2H), 2.40-1.99 (m, 6H), 1.45 (d, J=6.2 Hz, 3H), 1.03(s, 9H), 0.99 (s, 9H), 0.28 (s, 3H), 0.26 (s, 3H), 0.20 (s, 6H); transoxime ¹H NMR (CDCl₃, 400 MHz) δ 7.44 (bd, J=6.5 Hz, 2H), 7.37 (bdd,J=7.6, 6.9 Hz, 2H), 7.33-7.31 (m, 1H), 6.41 (s, 1H), 6.04-5.97 (m, 1H),5.48 (bd, J=15.0 Hz, 1H), 5.29-5.27 (m, 1H), 5.22 (bs, 2H), 5.00-4.95(m, 1H), 3.98-3.89 (m, 2H), 2.39-2.02 (m, 6H), 1.37 (d, J=5.9 Hz, 3H),1.04 (s, 9H), 0.99 (s, 9H), 0.28 (s, 3H), 0.27 (s, 3H), 0.23 (s, 3H),0.22 (s, 3H).

Example 22 General Procedure for the Synthesis of Compounds 2-158

As depicted in Scheme 21, to a solution of corresponding compound 2-157(1.0 equiv) in THF was added TBAF (2.5 equiv, 1M solution in THF) andthe mixture was stirred at room temperature for 2 hours. The solventswere then evaporated under reduced pressure, and filtration on SiO₂ witha mixture of 30% EtOAc/cyclohexane afforded after evaporation, compounds2-158 in >85% yield. An illustrative example of compound 2-158 follows;its characterization data are in the text below the figure.

Illustrative Compound 2-158:

Cis: ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 11.52 (s, 1H), 7.45-7.34 (m, 5H),6.64 (s, 1H), 6.09-6.02 (m, 2H), 5.34-5.25 (m, 4H), 5.18-5.08 (m, 2H),4.33 (d, J=17.0 Hz, 1H), 4.15 (d, J=17.6 Hz, 1H), 2.65-2.59 (m, 1H),2.27-2.14 (m, 3H), 2.04-2.00 (m, 1H), 1.88-1.83 (m, 1H), 1.30 (t, J=6.4Hz, 3H); HRMS (ESI) m/z 478.1372 ([M+Na⁺], C₂₅H₂₆ClNO₅Na requires478.1392).

Trans: ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 11.73 (s, 1H), 7.32-7.26 (m,5H), 6.64 (s, 1H), 6.50 (d, J=16.4 Hz, 1H), 6.06-5.98 (m, 2H), 5.43-5.24(m, 3H), 4.91 (s, 2H), 4.22 (s, 2H), 2.61-2.55 (m, 1H), 2.46-2.33 (m,2H), 2.20-2.02 (m, 3H), 0.98 (t, J=6.4 Hz, 3H); HRMS (ESI) m/z 478.1522([M+Na⁺], C₂₅H₂₆ClNO₅Na requires 478.1392).

Example 23 General Procedure for the Synthesis of Compounds 2a-1 viaMitsunobu Cyclization

As depicted in Scheme 25, the preparation of compound 2a-1 begins withcommercially available orcinol, 8. The description below is not intendedto be limiting and alternate analogs may be prepared with the samegeneral process.

Formylation of orcinol, synthesis of aldehyde 9. POCl₃ (54.9 mL, 600mmol, 2.0 equiv) was added slowly to a flask containing DMF (100 mL) at0° C. To this mixture was then added a solution of orcinol (42.65 g, 300mmol, 1.0 equiv) as a solution in DMF (100 mL) and the reaction wasallowed to warm up to 23° C. and stirred overnight. Then, the reactionwas cooled to 0° C. and 200 mL of ice-water were added. The pH of themixture was adjusted between 6 and 7 by addition of 10% of NaOH. Themixture was allowed to stand for 30 minutes and then the precipitateswere filtered to afford the desired aldehyde 9 (29.2 g, 64%) as a whitesolid which was used in the subsequent step without furtherpurification: Rf=0.31 (Hexane/EtOAc 3/1); ¹H NMR ((CD₃)₂CO, 400 MHz, 25°C.) δ 12.48 (s, 1H), 10.10 (s, 1H), 9.53 (s, 1H), 6.30 (d, J=2.4 Hz,1H), 6.17 (d, J=2.0 Hz, 1H), 2.54 (s, 3H); ¹³C NMR ((CD₃)₂CO, 100 MHz,25° C.) δ 194.3, 167.3, 166.2, 146.1, 113.8, 111.5, 101.4, 18.2; HRMS(MALDI-TOF) m/z 175.0373 ([M+Na⁺], C₈H₈O₃Na requires 175.0371).

Bis-EOM-protection, synthesis of aldehyde 9b. To a solution ofbis-phenol 9 (29.2 g, 191.9 mmol) in DMF (400 mL) at 0° C. weresequentially added iPr₂NEt (95.2 mL, 575.7 mmol, 3.0 equiv) and EOMCl(52.3 mL, 575.7 mmol, 3.0 equiv). The reaction was allowed to warm up to23° C. and stirred overnight. Then, the reaction was quenched with sat.NH₄Cl_(aq) (100 mL) and further diluted with CH₂Cl₂ (400 mL). Theorganic layer was separated and washed with sat. NH₄Cl_(aq) (100 mL×2),brine (100 mL×2), and dried over anhydrous Na₂SO₄ (5.0 g). Filtrationand evaporation of the solvents under reduced pressure followed by flashchromatography (SiO₂, Hexane/EtOAc 8/1) afforded protected aldehyde 9bas a colorless oil (46.5 g, 90%): Rf=0.51 (Hexane/EtOAc 3/1); ¹H NMR(CDCl₃, 400 MHz, 25° C.) δ 10.48 (s, 1H), 6.71 (d, J=2 Hz, 1H), 6.49 (d,J=2 Hz, 1H), 5.27 (s, 2H), 5.22 (s, 2H), 3.72 (q, J=6.9 Hz, 2H), 3.69(q, J=6.9 Hz, 2H), 2.54 (s, 3H), 1.20 (t, J=7.0 Hz, 3H), 1.19 (t, J=7.0Hz, 3H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 190.6, 163.0, 162.1, 144.2,118.6, 112.3, 100.6, 93.6, 92.8, 64.8, 64.7, 22.2, 15.1 (×2); HRMS(MALDI-TOF) m/z 291.1224 ([M+Na⁺], C₁₄H₂₀O₅Na requires 291.1209).

Oxidation-chlorination, synthesis of acid 10. To a solution of aldehyde9b (46.5 g, 173 mmol) in THF (200 mL) was added a solution of NaH₂PO₄(67.5 g, 433 mmol, 2.5 equiv) in H₂O (200 mL) and the mixture was cooleddown to 0° C. Then a solution of NaClO₂ (48.9 g, 85%, 433 mmol, 2.5equiv) in H₂O (200 mL) was added slowly to the reaction. After stirringovernight the mixture was diluted with EtOAc (500 mL), washed with brine(200 mL×3) and dried over Na₂SO₄ (5.0 g). Filtration and evaporation ofthe solvents under reduced pressure afforded acid 10 (49.6 g, 90%) as awhite solid which was used in the next step without furtherpurification: Rf=0.66 (CH₂Cl₂/MeOH 9/1); ¹H NMR ((CD₃)₂CO, 400 MHz, 25°C.) δ 7.08 (s, 1H), 5.34 (s, 2H), 5.25 (s, 2H), 3.75 (q, J=7.1 Hz, 2H),3.71 (q, J=7.1 Hz, 2H), 2.34 (s, 3H), 1.19 (t, J=7.1 Hz, 3H), 1.17 (t,J=7.1 Hz, 3H); ¹³C NMR ((CD₃)₂CO, 100 MHz, 25° C.) δ 168.1, 154.9,153.9, 135.1, 121.8, 117.3, 102.8, 94.7, 94.6, 65.2, 65.0, 17.6, 15.4,15.4; HRMS (MALDI-TOF) m/z 341.0713 ([M+Na⁺], C₁₄H₁₉ClO₆Na requires341.0768).

Mitsunobu esterification, synthesis of ester 11. To a solution of acid10 (23.5 g, 73.7 mmol) in toluene (300 mL) at 0° C., were addedsequentially trimethylsilylethanol (12.7 mL, 88.5 mmol, 1.2 equiv), PPh₃(28.9 g, 110.5 mmol, 1.25 equiv) and DIAD (21.7 mL, 110.5 mmol, 1.25equiv). The reaction was allowed to warm up to 23° C. and stirred for 3hours. Evaporation of the solvents under reduced pressure followed byflash chromatography (SiO₂, Hexane/EtOAc 20/1 and 10/1) afforded ester11 (24.7 g, 80%) as a white solid: Rf=0.60 (Hexane/EtOAc 3/1); ¹H NMR(CDCl₃, 400 MHz, 25° C.) δ 6.98 (s, 1H), 5.28 (s, 2H), 5.18 (s, 2H),4.41-4.36 (m, 2H), 3.75 (q, J=6.9 Hz, 2H), 3.70 (q, J=6.9 Hz, 2H), 2.32(s, 3H), 1.21 (t, J=7.1 Hz, 3H), 1.20 (t, J=7.1 Hz, 3H), 1.12-1.07 (m,2H), 0.06 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 167.7, 154.2,153.1, 135.0, 120.5, 117.3, 102.0, 94.0, 93.9, 64.7, 64.5, 63.8, 17.6,15.1 (×2), −1.41 (×3); HRMS (MALDI-TOF) m/z 441.1487 ([M+Na⁺],C₁₉H₃₁ClO₆SiNa requires 441.1476).

Reduction of diester 16, synthesis of diol 17a. To the suspension ofLiAlH₄ (15.2 g, 400 mmol, 4 equiv) in THF (300 mL) at 0° C. was addeddrop wise a solution of trans-3-hexenedioic acid dimethyl ester (17.2 g,100 mmol) in THF (100 mL). The reaction was stirred for 2 hours. Afterwhich, the reaction was quenched by addition of H₂O (15.2 mL), 15% ofNaOH (15.2 mL), and H₂O again (45.6 mL). Then the mixture was pouredinto Et₂O (150 mL), stirred for 30 minutes, and filtered over Na₂SO₄ (5g). Evaporation of the solvents under reduced pressure afforded diol 17awhich was used without further purification (11.0 g, 95%): Rf=0.53(CH₂Cl₂/MeOH 9/1); ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 5.55-5.52 (m, 2H),3.66 (dt, J=5.6, 5.2 Hz, 2H), 2.33-2.28 (m, 4H), the OH is not visible;¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 129.8 (×2), 62.0 (×2), 36.16 (×2);HRMS (MALDI-TOF) m/z 139.0734 ([M+Na⁺], C₆H₁₂O₂Na requires 139.0735).

TBDPS-monoprotection of 17a, synthesis of alcohol 17. To a suspension ofNaH (1.90 g, 47.4 mmol, 1.0 equiv) in THF (100 mL) at 0° C. a solutionof diol 17a (5.5 g, 47.4 mmol, 1.0 equiv) in THF (20 mL) was added.After stirring for 45 minutes, TBDPSCl (12.4 ml, 47.4 mmol, 1.0 equiv)was added. The reaction mixture was allowed to warm up to 23° C. andstirred for 1 hour. Then, it was quenched with sat. NH₄Cl_(aq) (200 mL)and extracted with EtOAc (150 mL×3), the combined organic layers werewashed with brine (200 mL×2) and dried over Na₂SO₄ (10 g). Filtrationand evaporation of the solvents under reduced pressure followed by flashchromatography (SiO₂, Hexane/EtOAc 20/1, 10/1 and 5/1) afforded desiredalcohol 17 (11.5 g, 68%) as a colorless oil: Rf=0.46 (Hexane/EtOAc 3/1);¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.68-7.65 (m, 4H), 7.43-7.36 (m, 6H),5.56 (dt, J=15.2, 6.4 Hz, 1H), 5.44 (dt, J=15.2, 6.8 Hz, 1H), 3.69 (t,J=6.4 Hz, 2H), 3.62 (t, J=6.3 Hz, 2H), 2.31-2.24 (m, 4H), 1.05 (s, 9H),the OH is not visible; HRMS (MALDI-TOF) m/z 377.1927 ([M+Na⁺],C₂₂H₃₀O₂SiNa requires 377.1913).

Synthesis of nitrile 19. To a solution of alcohol 17 (11.5 g, 32.4 mmol)in CH₂Cl₂ (150 mL) at 0° C. were added imidazole (4.30 g, 47.0 mmol,1.45 equiv), triphenylphosphine (9.35 g, 35.7 mmol, 1.1 equi) and iodine(9.06 g, 35.7 mmol, 1.1 equiv) and the resulting solution was stirred at0° C. for 6 hours. Then, the mixture was diluted with Et₂O/Hexane 1/1(300 mL), washed with sat. NaHCO_(3aq) (150 mL×2), sat. Na₂S₂O_(5aq)(150 mL), brine (150 mL) and dried over Na₂SO₄ (5.0 g). The solventswere evaporated and resulting oily solids were triturated with hexanes(200 mL×2). Filtration and evaporation of the hexanes afforded thecorresponding alkyl iodide 18 as a colorless oil which was directly usedin the next step without further purification. Rf=0.46 (Hexane/EtOAc3/1); ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.70-7.67 (m, 4H), 7.45-7.36 (m,6H), 5.58 (dt, J=15.2, 6.8 Hz, 1H), 5.44-5.37 (dt, J=15.2, 6.4 Hz, 1H),3.69 (t, J=6.4 Hz, 2H), 3.11 (t, J=7.2 Hz, 2H), 2.54 (dt, J=7.2, 6.8 Hz,2H), 2.26 (dt, J=6.4, 6.4 Hz, 2H), 1.05 (s, 9H); HRMS (MALDI-TOF) m/z487.0923 ([M+H⁺], C₂₂H₂₉IOSiH requires 487.0930).

To a solution of crude alkyl iodine 18 in DMSO (200 mL) was added KCN(21.0 g, 324 mmol, 1.0 equiv). The resulting mixture was stirred for 2hours at 60° C. and then the reaction mixture was cooled down to 23° C.,diluted with EtOAc (200 mL), washed with water (200 mL), brine (100 mL),and dried over Na₂SO₄ (5.0 g). Filtration and evaporation of thesolvents under reduced pressure followed by flash chromatography (SiO₂,Hexane/EtOAc 20/1 and 10/1) afforded desired nitrile 19 (10.6 g, 90%) asa colorless oil: Rf=0.46 (Hexane/EtOAc 3/1); ¹H NMR (CDCl₃, 400 MHz, 25°C.) δ 7.71-7.69 (m, 4H), 7.45-7.39 (m, 6H), 5.60 (dt, J=15.2, 6.4 Hz,1H), 5.52-545 (m, 1H), 3.73 (t, J=6.4 Hz, 2H), 2.36-2.28 (m, 6H), 1.08(s, 9H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.) 135.7 (×4), 134.0 (×2), 130.5,129.7 (×2), 127.8, 127.7 (×4), 119.4, 63.6, 35.9, 28.6, 27.0 (×3), 19.3,17.6; HRMS (MALDI-TOF) m/z 386.1962 ([M+Na⁺], C₂₃H₂₉OSiNNa requires386.1916).

Synthesis of ylide 21. To a solution of N-methoxy-N-methylhydroxylaminehydrochloride (48.52 g, 500 mmol, 1.0 equiv) in CH₂Cl₂ (1.0 L) at 0° C.were added pyridine (80.63 mL, 1.0 mol) and chloroacetic anhydride(85.14 g, 500 mmol, 1.0 equiv). The resulting mixture was stirred for 15min at 0° C., then warm up to 23° C. and stirred overnight. The reactionmixture was then poured carefully into sat. NaHCO_(3aq) solution (1.0 L)and stirred 1 hour, after which the layers were separated, the aqueousphase was extracted with CH₂Cl₂ (400 mL) and the combined organic layerswere washed with 1N HCl (200 mL×2), brine (200 mL×2), dried over Na₂SO₄(10 g) and filtered. Evaporation of the solvents under reduced pressureafforded the corresponding acetamide (N-methoxy-N-methylacetamide-2-chloride) as a green oil which was used in the next stepwithout further purification. To a solution of this acetamide in CH₃CN(800 mL) was added Ph₃P (107.98 g, 411.7 mmol, 0.82 equiv) and theresulting mixture was refluxed for 18 hours. Then the solvents wereremoved under vacuum and the resulting viscous oil was dissolved inCH₂Cl₂ (1.0 L), washed with 2N KOH (400 mL×2), brine (400 mL) and driedover Na₂SO₄ (10.0 g). Filtration and evaporation of the solvents underreduced pressure afforded ylide 21 as a thick oil which solidified bystanding (146.5 g, 80% over two steps). This compound was used in thenext step without further purification. Rf=0.85 (Hexane/EtOAc 3/1); ¹HNMR (CDCl₃, 400 MHz, 25° C.) δ 7.71-7.65 (m, 6H), 7.55-7.50 (m, 3H),7.48-7.42 (m, 6H), 3.74 (s, 3H), 3.08 (s, 3H), 1.86 (s, 1H); ¹³C NMR(CDCl₃, 100 MHz, 25° C.) 133.3 (×3), 133.2 (×3), 131.9 (×3), 128.9 (×3),128.8 (×3), 127.9, 61.3, 35.9.

Synthesis of Weinreb amide 12. To a solution of nitrile 19 (22.94 g,63.0 mmol, 1.0 equiv) in CH₂Cl₂ (250 mL) at −78° C., DIBAL (66.3 mL, 1Min toluene, 1.05 equiv) was added and the reaction was stirred for 30min. To quench the reaction, a solution of sat. K/Na(tartrate)_(aq) (300mL) was added and the biphasic mixture was stirred until it became aclear biphasic system (over 2 h). The two phases were separated and theaqueous phase was further extracted with CH₂Cl₂ (200 mL×3). The combinedorganic layers were then washed with brine (200 mL) and dried overNa₂SO₄ (10 g). Filtration followed by evaporation of the solvents underreduced pressure afforded the corresponding aldehyde 20 which was usedin the next step without further purification. Thus, the crude aldehyde20 was dissolved in CH₂Cl₂ (200 mL) at 23° C. and ylide 21 was added (30g, 81.9 mmol, 1.3 equiv). The reaction was then stirred overnight.Evaporation of the solvents under reduced pressure followed by flashchromatography (SiO₂, Hexane/EtOAc 20/1, 10/1 and 3/1) afforded desiredWeinreb amide 12 (14.5 g, 61% over two steps) as a colorless oil:Rf=0.30 (Hexane/EtOAc 3/1); ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.69-7.67(m, 4H), 7.43-7.36 (m, 6H), 6.98 (dt, J=15.3, 7.1 Hz, 1H), 6.40 (d,J=15.3 Hz, 1H), 5.48-5.46 (m, 2H), 3.68 (t, J=6.6 Hz, 2H), 3.67 (s, 3H),3.24 (s, 3H), 2.32-2.24 (m, 4H), 2.19-2.14 (m, 2H), 1.06 (s, 9H); ¹³CNMR (CDCl₃, 100 MHz, 25° C.) 167.1, 147.2, 135.7 (×4), 134.1, 131.0(×2), 129.6 (×2), 127.8 (×4), 127.7, 119.0, 64.0, 61.7, 36.1, 32.6,32.5, 31.5, 27.0 (×3), 19.3; HRMS (MALDI-TOF) m/z 474.2432 ([M+Na⁺],C₂₇H₃₇NO₃SiNa requires 474.2440).

Coupling of toluate 11 and Weinreb amide 12, synthesis of ketone 13. Asolution of compound 11 (8.38 g, 20.0 mmol, 1.0 equiv) in anhydrous THF(120 mL) at −78° C. was treated with freshly prepared LDA (71.4 mL, 0.56M, 40.0 mmol, 2.0 equiv) added via cannula. Immediately after, asolution of Weinreb amide 12 (8.13 g, 18 mmol, 0.9 equiv) in THF (15 mL)at −78° C. was added via cannula. The resulting mixture was then stirredfor 15 minutes and the reaction was quenched by addition of sat.NH₄Cl_(aq) (20 mL). Upon warming to 23° C., the reaction mixture wasextracted with EtOAc (200 mL×2), and the combined organic layers werethen washed with brine (100 mL) and dried over Na₂SO₄ (5.0 g).Filtration and evaporation of the solvents under reduced pressurefollowed by flash chromatography (SiO₂, Hexane/EtOAc 20/1, 10/1 and 5/1)afforded desired ketone 13 (20.7 g, 65%): Rf=0.48 (Hexane/EtOAc 3/1); ¹HNMR (CDCl₃, 400 MHz, 25° C.) δ 7.70-7.68 (m, 4H), 7.42-7.38 (m, 6H),7.10 (s, 1H), 6.94 (dt, J=15.8, 6.7 Hz, 1H), 6.19 (d, J=15.8 Hz, 1H),5.49-5.46 (m, 2H), 5.30 (s, 2H), 5.22 (s, 2H), 4.36-4.32 (m, 2H), 4.06(s, 2H), 3.79-3.68 (m, 6H), 2.30-2.25 (m, 4H), 2.18-2.13 (m, 2H), 1.23(t×2, J=7.1 Hz, 6H), 1.07 (s, 9H), 1.08-1.03 (m, 2H), 0.06 (s, 9H); ¹³CNMR (CDCl₃, 100 MHz, 25° C.) δ 194.5, 167.0, 154.6, 153.9, 147.1, 135.5(×4), 133.9 (×2), 132.8, 130.5, 129.5 (×2), 129.1, 127.9, 127.6 (×4),120.3, 117.7, 103.1, 93.9, 93.8, 64.6, 64.4, 63.8, 63.6, 60.3, 59.9,43.0, 35.9, 32.4, 31.1, 26.8 (×3), 22.6, 22.0, 19.2, 17.4, 15.0, 14.2,14.1, −1.40 (×3); HRMS (MALDI-TOF) m/z 831.3380 ([M+Na⁺],C₄₄H₆₁ClO₈Si₂Na requires 831.3491).

Synthesis of oximes 14a. To a solution of ketone 13 (18.6 g, 25.5 mmol)in pyridine (50 mL) at 40° C., carboxymethoxylamine hemihydrochloride(13.98 g, 127.5 mmol) was added and the reaction was stirred at suchtemperature for 24 hours. After evaporation of the pyridine, theresidues were dissolved in CH₂Cl₂ (200 mL) and washed with sat.NH₄Cl_(aq) (50 mL×2), brine (50 mL×2) and dried over anhydrous Na₂SO₄(2.0 g). Filtration and evaporation of the solvents under reducedpressure followed by flash chromatography (SiO₂, Hexane/EtOAc 5/1 and1/1) afforded desired oximes 14a (10.1 g, 50%) as a mixture of E/Zisomers in a 1/1 ratio: Rf=0.51 (CH₂Cl₂/MeOH 19/1); ¹H NMR (CDCl₃, 400MHz, 25° C.) δ 7.69-7.65 (m, 8H), 7.42-7.35 (m, 12H), 7.07 (s, 1H), 7.05(s, 1H), 6.74 (d, J=15.8 Hz, 1H), 6.36 (dt, J=15.8, 6.8 Hz, 1H), 6.07(dt, J=15.8, 6.8 Hz, 1H), 5.79 (d, J=15.8 Hz, 1H), 5.48-5.43 (m, 2H),5.39-5.34 (m, 2H), 5.30 (s, 2H), 5.28 (s, 2H), 5.21 (s, 2H), 5.19 (s,2H), 4.63 (s, 2H), 4.46 (s, 2H), 4.35-4.29 (m, 4H), 4.03 (s, 2H), 3.88(s, 2H), 3.80-3.60 (m, 12H), 2.31-2.25 (m, 6H), 2.17-2.15 (m, 2H),2.10-2.06 (m, 2H), 2.00-1.97 (m, 2H), 1.27-1.21 (m, 12H), 1.08 (s, 9H),1.07 (s, 9H), 1.07-1.05 (m, 4H), 0.09 (s, 9H), 0.08 (s, 9H), the OH fromthe acids are not visible; ¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 172.0,171.9, 167.9, 167.3, 157.0, 155.6, 154.6, 154.5, 153.7, 153.7, 141.4,137.8, 135.7 (×8), 134.6, 134.1 (×4), 133.7, 131.2, 130.8, 129.7 (×4),127.9, 127.7 (×8), 127.5, 124.2 (×2), 120.4, 119.1, 117.5, 117.4, 103.2,102.8, 94.1, 94.0, 93.9, 93.8, 71.1, 70.5, 64.9, 64.9, 64.7, 64.6, 64.5,64.0, 64.0, 63.9, 36.1 (×2), 33.5, 33.1, 32.9, 31.9, 31.9, 29.4, 27.0(×6), 19.3 (×2), 17.5, 17.3, 15.1 (×4), −1.4 (×6).

Synthesis of amides 14. To the solution of acid 14a (10.0 g, 11.33 mmol,1.0 equiv) in CH₂Cl₂ (60 mL) at 0° C., was added EDC.HCl (2.6 g, 13.6mmol, 1.2 equiv) followed by a solution of piperidine (1.34 mL, 13.6mmol, 1.2 equiv) in CH₂Cl₂ (10 mL). The reaction was allowed to warm upto 23° C., and stirred for 3 hours. The reaction mixture was thendiluted with CH₂Cl₂ (200 mL), washed with sat. NH₄Cl_(aq) (100 mL),brine (100 mL), and dried over anhydrous Na₂SO₄ (5 g). Filtration andevaporation of the solvents under reduced pressure followed by flashchromatography (SiO₂, Hexane/EtOAc 2/1 and 1/1) afforded desired amides14 (6.60 g, 63%) as a mixture of two isomers in a 1/1 ratio: Rf=0.31(Hexane/EtOAc 3/1); ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.68-7.64 (m, 8H),7.41-7.33 (m, 12H), 7.04 (s×2, 2H), 6.71 (d, J=16.0 Hz, 1H), 6.21 (dt,J=16.0, 6.7 Hz, 1H), 6.07 (dt, J=16.0, 6.7 Hz, 1H), 5.73 (d, J=16.0 Hz,1H), 5.45-5.39 (m, 2H), 5.37-5.32 (m, 2H), 5.28 (s, 2H), 5.25 (s, 2H),5.19 (s, 4H), 4.73 (s, 2H), 4.60 (s, 2H), 4.37-4.27 (m, 4H), 3.95 (s,2H), 3.87 (s, 2H), 3.80-3.60 (m, 12H), 3.55-3.52 (m, 2H), 3.51-3.48 (m,2H), 3.38-3.35 (m, 2H), 3.32-3.29 (m, 2H), 2.26-2.16 (m, 6H), 2.09-2.06(m, 2H), 2.01-1.98 (m, 2H), 1.95-1.91 (m, 2H), 1.61-1.46 (m, 12H),1.28-1.17 (m, 12H), 1.05-1.01 (m, 4H), 1.04 (s, 9H), 1.03 (s, 9H), 0.05(s, 9H), 0.04 (s, 9H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 167.1, 167.0(×2), 166.8, 155.6, 154.3, 154.2, 153.5, 153.4, 153.1, 139.2, 136.5,135.6 (×8), 134.7, 134.6, 134.1 (×4), 131.4, 131.1, 129.6 (×4), 127.6(×8), 127.6, 127.1, 124.1, 121.0, 120.9, 119.5, 117.9, 117.7, 103.1,102.9, 94.1, 94.0, 93.9, 73.1, 73.0, 64.8, 64.7, 64.6, 64.5, 64.0, 63.9,63.9, 63.6, 60.4, 53.5, 46.4, 46.2, 43.0, 42.9, 36.1, 33.4, 33.0, 32.9,32.0, 31.9, 30.0, 26.9 (×6), 26.6, 26.5, 25.6, 24.6, 19.3 (×2), 17.4,17.3, 15.1 (×6), −1.42 (×6).

Silyl deprotection, synthesis of macrocyclization precursor 14b. To asolution of ester 14 (6.60 g, 6.9 mmol, 1.0 equiv) in THF (70 mL) at 23°C. TBAF (17.3 mL, 1M in THF, 2.5 equiv) was added dropwise and theresulting mixture was stirred for 3 hours. Then, EtOAc (100 mL) wasadded to the reaction, and the resulting mixture was washed with 1M HCl(60 mL×3), brine (60 mL), and dried over Na₂SO₄ (1.0 g). Filtration andevaporation of the solvents under reduced pressure followed by flashchromatography (SiO₂, EtOAc and EtOAc/MeOH 4/1) afforded desired acids14b (3.78 g, 89%) as a mixture of two isomers in a 1/1 proportion:Rf=0.35 (CH₂Cl₂/MeOH 9/1); ¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 7.03 (s,1H), 6.99 (s, 1H), 6.62 (d, J=16.1 Hz, 1H), 6.23 (dt, J=16.1, 6.9 Hz,1H), 6.11 (dt, J=16.1, 6.9 Hz, 1H), 5.95 (d, J=16.1 Hz, 1H), 5.55-5.30(m, 4H), 5.26 (s×2, 4H), 5.21 (s, 2H), 5.20 (s, 2H), 4.72 (s, 2H), 4.63(s, 2H), 4.05 (s, 2H), 4.00 (s, 2H), 3.81-3.66 (m, 8H), 3.65-3.57 (m,4H), 3.54 (t, J=5.4 Hz, 2H), 3.44 (t, J=5.4 Hz, 2H), 3.31 (t, J=5.4 Hz,2H), 3.24 (t, J=5.4 Hz, 2H), 2.23 (dt, J=6.5, 6.5 Hz, 4H), 2.18-2.04 (m,8H), 1.67-1.49 (m, 10H), 1.42-1.40 (m, 2H), 1.25-1.16 (m, 12H), OH fromthe alcohols and the acids are not visible; ¹³C NMR (CDCl₃, 100 MHz, 25°C.) δ 168.73, 168.03, 156.47, 154.19, 152.99, 152.94, 140.23, 136.77,134.37, 134.18, 132.45, 132.11, 127.53, 127.23, 125.91, 122.46, 119.28,116.76, 102.91, 102.80, 94.16, 94.02, 93.96, 71.51, 71.25, 64.92, 64.82,64.54, 64.46, 62.11, 46.24, 45.60, 43.39, 43.27, 36.06, 35.98, 33.08,32.80, 32.09, 31.86, 31.61, 29.20, 26.26, 26.18, 25.38, 25.30, 24.41,24.30, 15.28, 15.16; HRMS (MALDI-TOF) m/z 633.2562 ([M+Na⁺]C₃₀H₄₃ClN₂O₉Na requires 633.2555).

Macrocyclization of 14b, synthesis of compounds 15. To a solution ofacid 14b (3.78 g, 6.18 mmol) in toluene (200 mL) at 0° C., was addedPPh₃ (2.43 g, 9.27 mmol) followed by a slow addition of DIAD (1.83 mL,9.3 mmol). The reaction was allowed to warm to 23° C. and stirred for 5hours. Evaporation of the solvents under reduced pressure followed byflash chromatography (SiO₂, Hexane/EtOAc 4/1 and 1/1) afforded desiredmacrocycles 15 (2.09 g, 60%) as a mixture of two isomers in a 1/1proportion: Rf=0.50 (Hexane/EtOAc 1/1); ¹H NMR (CDCl₃, 400 MHz, 25° C.)δ 7.05 (s, 1H), 7.03 (s, 1H), 6.51 (d, J=16.0 Hz, 1H), 6.16 (dt, J=15.8,6.5 Hz, 1H), 6.00 (dt, J=15.8, 7.7 Hz, 1H), 5.44 (d, J=16.0 Hz, 1H),5.32 (m, 4H), 5.28 (s, 2H), 5.26 (s, 2H), 5.21 (s, 2H), 5.20 (s, 2H),4.80 (s, 2H), 4.71 (s, 2H), 4.23 (t, J=5.1 Hz, 2H), 4.18 (t, J=5.1 Hz,2H), 3.91 (bs, 2H), 3.80-3.66 (m, 10H), 3.60-3.51 (m, 4H), 3.48-3.46 (m,2H), 3.41-3.38 (m, 2H), 2.40-2.32 (m, 4H), 2.19-2.16 (m, 1H), 2.11-2.00(m, 7H), 1.67-1.51 (m, 12H), 1.23-1.19 (12H); ¹³C NMR (CDCl₃, 100 MHz,25° C.) δ 167.5, 166.9, 154.8, 154.6, 154.4, 153.2, 141.0, 136.6, 134.1,132.6, 128.5, 127.6; 124.6, 121.6, 119.5, 117.5, 102.94 (×2), 94.1 (×2),93.8 (×2), 73.4, 73.2, 72.8, 72.7, 64.9, 64.8, 64.7 (×2), 64.3 (×2),46.5, 46.3, 43.1 (×2), 35.7, 35.6, 32.5, 32.4, 32.2, 32.1, 31.9, 31.7,30.2 (×2), 26.7 (×2), 26.6 (×2), 25.7 (×2), 24.7 (×4), 15.2 (×2), 15.1(×2); HRMS (MALDI-TOF) m/z 615.2438 ([M+Na⁺], C₃₀H₄₁ClN₂O₈Na requires615.2450).

Deprotection, synthesis of 2a-1 as an E/Z mixture, isolation of the Eisomer and isomerization of the Z isomer. To a solution of macrocycles15 (2.8 g, 4.72 mmol) in MeOH (60 mL) at 40° C. sulfonic acid resin(7.87 g, 3.0 mmol/g, 23.6 mmol) was added, and the suspension wasstirred for 2 hours. The mixture was diluted with CH₂Cl₂ (60 mL),filtered, and the resin was once rinsed with CH₂Cl₂ (20 mL×2). Afterremoval of the solvent, the residue was re-dissolved in MeOH (50 mL) andsonicated. A precipitate began to form and the solution was allowed tostand for 12 h. The solution was filtered and rinsed with MeOH (30 mL).Evaporation of the combined solutions afforded 2a-1 (1.13 g, 9:1 E:Z asjudged by LCMS). The remaining solid (pure Z-isomer, 900 mg) wasdissolved in CH₂Cl₂ (180 mL) and treated with TFA (1.4 mL, 18.8 mmol) at23° C. The mixture was stirred for 12 hours, after which toluene (50 mL)was added and the solvents were evaporated (LCMS of this crude indicateda 1:1 mixture of E/Z isomers). The residue obtained was re-dissolved inCH₂Cl₂ (30 mL). The insoluble precipitated was filtered and rinsed withMeOH (20 mL). Evaporation of the combined solutions afforded more ofcompound 2a-1 (450 mg) again as a 9:1 mixture in favor of the desired Eisomer. The remaining solid (Z-isomer, 450 mg) was submitted to sameisomerization conditions two more times. The combined batches of 8 werethen purified by flash chromatography (70 g of C18, CH₃CN/H₂O 35/65 and0.01% TFA, 50 mL/min) to afford the pure E isomer as a white powder(1.10 g, 51% overall yield); Rf=0.44 (Hexane/EtOAc 1/2); ¹H NMR (CDCl₃,400 MHz, 25° C.) δ 11.64 (s, 1H), 6.64 (s, 1H), 6.01 (td, J=15.5 7.5 Hz,1H), 5.11 (d, J=15.5 Hz, 1H), 5.10-5.03 (m, 2H), 4.85 (s, 2H), 4.37 (t,J=4.8 Hz, 2H), 4.17 (s, 2H), 3.60 (t, J=5.0 Hz, 2H), 3.46 (t, J=5.0 Hz,2H), 2.34 (q, J=5.4 Hz, 2H), 2.10-2.02 (m, 2H), 1.99-1.92 (m, 2H),1.70-1.54 (m, 6H); ¹³C NMR (CDCl₃, 100 MHz, 25° C.) δ 170.27, 167.42,163.21, 157.38, 155.18, 138.21, 135.62, 131.82, 129.13, 124.76, 115.55,107.60, 103.47, 72.63, 65.03, 46.38, 43.31, 33.21, 32.76, 31.94, 31.84,26.65, 25.64, 24.57; HRMS (MALDI-TOF) m/z 499.1638 ([M+H⁺],C₂₄H₂₉ClN₂O₆H requires 499.1612).

Compound a2-13:

¹H NMR (CDCl₃, 400 MHz, 25° C.) δ 11.65 (s, 1H), 6.86 (d, J=1.8 Hz, 1H),6.33 (d, J=1.8 Hz, 1H), 6.14 (td, J=15.8, 7.6 Hz, 1H), 5.82 (d, J=15.7Hz, 1H), 5.33 (bs, 2H), 4.85 (s, 2H), 4.53 (bs, 2H), 4.33 (s, 2H), 3.57(t, J=5.0 Hz, 2H), 3.39 (t, J=5.0 Hz, 2H), 2.50 (bs, 2H), 2.11-2.06 (m,4H), 1.66-1.58 (m, 6H); ¹³C NMR (DMSO-d₆, 100 MHz, 25° C.) δ 168.5,166.3, 159.3, 157.1, 156.2, 137.6, 136.0, 131.5, 129.5, 124.9, 113.1,105.9, 100.9, 71.9, 63.7, 45.2, 42.1, 32.3, 31.8, 30.5, 28.1, 26.0,25.3, 24.0; HRMS (MALDI-TOF) m/z 465.2054 ([M+Na⁺], C₂₄H₃₀N₂O₆H requires465.2002).

Example 24 Kinase Inhibition of Exemplary Compounds

A representative subset of the pochonin analogue library (84 compounds)was tested for its inhibition in a panel of 24 kinase (AKT1, ARK5,Aurora-A, Aurora-B, B-RAF-VE, CDK2/CycA, CDK4/CycD1, CK2-α1, FAK, EPHB4,ERB2, EGF-R, IGF1-R, SRC, VEGF-R2, VEGF-R3, FLT3, INS-R, MET, PDGFR-β,PLK1, SAK, TIE2, COT) at 10 μM using the procedure described below.

All protein kinases were expressed in Sf9 insect cells as humanrecombinant GST-fusion proteins or His-tagged proteins by means of thebaculovirus expression system. Kinases were purified by affinitychromatography using either GSH-agarose (Sigma) or Ni-NTH-agarose(Qiagen). The purity of each kinase was checked by SDS-PAGE/silverstaining and the identity of each kinase was verified by western blotanalysis with kinase specific antibodies or by mass spectroscopy.

Protein Kinase Assay

A radiometric protein kinase assay (³³PanQinase® Activity Assay) wasused for measuring the kinase activity of the 24 protein kinases. Allkinase assays were performed in 96-well FlashPlates™ from Perkin Elmer(Boston, Mass., USA) in a 50 μl reaction volume. The reaction cocktailwas pipetted in 4 steps in the following order:

-   -   20 μl of assay buffer    -   5 μl of ATP solution (in H₂O)    -   5 μl of test compound (in 10% DMSO)    -   10 μl of substrate/10 μl of enzyme solution (premixed)

The assay for all enzymes contained 60 mM HEPES-NaOH, pH 7.5, 3 mMMgCl₂, 3 mM MnCl₂, 3 μM Na-orthovanadate, 1.2 mM DTT, 50 μg/ml PEG₂₀₀₀₀,1 μM [γ-³³P]-ATP (approx. 5×10⁰⁵ cpm per well).

For the 24 kinase assays, the following amounts of enzyme and substratewere used per well:

Kinase Kinase Substrate # Kinase Lot # ng/50 μl Substrate ng/50 μl 1AKT1 SP007 100 GSK3(14-27), Lot 005 1000 2 ARK5 002 100 Autosphos. — 3Aurora-A SP004 50 tetra(LRRWSLG) 500 4 Aurora-B SP007 100 tetra(LRRWSLG)250 5 B-RAF-VE 001 20 MEK1-KM(Lot 013) 250 6 CDK2/CycA SP005 100 HistoneH1 125 7 CDK4/CycD1 006 50 Rb-CTF, Lot 010 500 8 COT 017 400Autophosphorylation — 9 EGF-R SP014 25 Poly(Glu,Tyr)_(4:1) 125 10 EPHB4SP006 10 Poly(Glu,Tyr)_(4:1) 125 11 ERBB2 SP011 200 Poly(Glu,Tyr)_(4:1)125 12 FAK SP006 100 Poly(Glu,Tyr)_(4:1) 125 13 IGF1-R 012 20Poly(Glu,Tyr)_(4:1) 125 14 SRC 004 10 Poly(Glu,Tyr)_(4:1) 125 15 VEGF-R2011 50 Poly(Glu,Tyr)_(4:1) 125 16 VEGF-R3 SP011 100 Poly(Glu,Tyr)_(4:1)125 17 FLT3 SP007 100 Poly(Ala,Glu,Lys,Tyr)_(6:2:5:1) 125 18 INS-R SP00525 Poly(Ala,Glu,Lys,Tyr)_(6:2:5:1) 125 19 MET SP011 100Poly(Ala,Glu,Lys,Tyr)_(6:2:5:1) 125 20 PDGFR-beta SP012 50Poly(Ala,Glu,Lys,Tyr)_(6:2:5:1) 125 21 PLK1 007 50 Casein 250 22 SAK 002200 Autosphosphorylation — 23 TIE2 SP006 200 Poly(Glu,Tyr)_(4:1) 250 24CK2-alpha1 SP003 200 Casein 1000

The reaction cocktails were incubated at 30° C. for 80 minutes. Thereaction was stopped with 50 μl of 2% (v/v) H₃PO₄, plates were aspiratedand washed two times with 200 μl of 0.9% (w/v) NaCl or 200 μl H₂O.Incorporation of ³³P_(i) was determined with a microplate scintillationcounter (Microbeta Trilux, Wallac).

All assays were performed with a BeckmanCoulter/Sagian robotic system.

Evaluation of Raw Data

The median value of the counts in column 1 (n=8) of each assay plate wasdefined as “low control”. This value reflects unspecific binding ofradioactivity to the plate in the absence of a protein kinase but in thepresence of the substrate. The median value of the counts in column 7 ofeach assay plate (n=8) was taken as the “high control”, i.e. fullactivity in the absence of any inhibitor. The difference between highand low control was taken as 100% activity.

As part of the data evaluation the low control value from a particularplate was subtracted from the high control value as well as from all 80“compound values” of the corresponding plate. The residual activity (in%) for each well of a particular plate was calculated by using thefollowing formula:

Res. Activity (%)=100×[(cpm of compound−low control)/(high control−lowcontrol)]

The residual activities for each concentration and the compound IC₅₀values were calculated using Quattro Workflow V2.1.0.0 (Quattro ResearchGmbH, Munich, Germany; www.quattro-research.com). The model used was“Sigmoidal response (variable slope)” with parameters “top” fixed at100% and “bottom” at 0%.

Significantly, twelve compounds showed more than 50% inhibition, whichrepresents a >14% hit rate for a kinase. Surprisingly, pochonin D,pochonin A and radicicol, though they had been shown to be powerfulinhibitors of HSP90, showed no significant activity against this panelof kinases. Nine compounds were selected to calculate IC₅₀ against eachof the 24 kinases (table 4). In this more detailed analysis, radicicolshowed only very mild activity against VGFR-R2 with no inhibition forthe twenty-three other kinases. Several pochonin analogues showed awell-defined pattern of activity against therapeutically relevantenzymes such as Src (8 μM for A2), Aurora A (12 μM for A3), IGF1-R (11μM for A5). Importantly, the compounds that proved to be kinaseinhibitors were not inhibitors of HSP90 and are not indiscriminateATP-surrogates.

Another subset of the library was tested for HSP90 inhibition bymeasuring direct interaction in a competitive assay and measuringdepletion of HSP90 client proteins in a cellular assay. HSP90's ATPasepocket has a specific fold that is present in a superfamily whichincludes functionally diverse proteins such as DNA topoisomerase II,helicase, MutL and histidine kinases (Bergerat fold). (A. Bergerat etal., Nature, 386:414 (1997); R. Dutta and M. Inouye, Trends Biochem.Sci., 25:24 (2000)). In fact, it has been shown that radicicol doesinhibit other members of this family albeit with lower affinity. (D.Gadelle et al., Nucleic Acids Res., 33:2310 (2005); P. G. Besant et al.,Mol. Pharmacol., 62:289 (2002). Yet remarkably, the best HSP90inhibitors of the invention were selective for HSP90 with respect to thepanel of kinases. Sixteen compounds were found to have an IC50<1 μM.

TABLE 4 Inhibitory activity (IC₅₀: μM) of selected pochonin analogs in apanel of 24 kinase assays (a blank represents an IC₅₀ >50 μM). Radi-Resorcylic Acid Species Kinase cicol A1 A2 A3 A4 A5 A6 A7 A8 AKT1 ARK5Aurora-A 14 12 30 47 Aurora-B 16 14 36 16 B-RAF-VE 50 CDK2/CycACDK4/CycD1 50 30 48 45 37 CK2-α1 FAK 14 9 37 38 34 EPHB4 16 40 49 40ERBB2 16 24 EGF-R 33 10 14 22 16 32 IGF1-R 23 16 19 13 11 21 SRC 11 8 1412 12 20 VEGF-R2 49 19 20 30 19 25 VEGF-R3 40 19 31 31 34 FLT3 23 23 4544 INS-R 36 44 MET 32 29 36 PDGFR-β PLK1 SAK 17 25 19 20 17 TIE2 72 1615 25 COT

In this detailed analysis, radicicol showed only very mild activityagainst VGFR-R2 with no inhibition for the twenty three other kinases.Several pochonin analogs showed a well-defined pattern of activityagainst therapeutically relevant enzymes, such as Src (8 μM for A2),Aurora A (12 μM for A3), and IGF1-R (11 μM for A5). Importantly, thecompounds that were found to be kinase inhibitors were not inhibitors ofHSP90 (data not shown) and are not indiscriminate ATP-surrogates. TheATP-binding pocket of HSP90 targeted by radicicol and pochonin D has aspecific fold that is present in a superfamily which includesfunctionally diverse proteins, such as DNA topoisomerase II, helicase,MutL, and histidine kinases (R. Dutta and M. Inouye, Trends Biochem.Sci., 24:24 (2000)). In fact, it has been shown that radicicol doesinhibit other members of this family albeit with lower affinity (D.Gadelle et al., Nucleic Acids Res., 33:2310 (2005); P. G. Besant et al.,Mol. Pharmacol., 62:289 (2002)). While the pochonin library describedabove will certainly contain some compounds that are good inhibitors ofenzymes bearing a Bergerat fold, we wished to evaluate whethermodification around the pochonin scaffold could retune the selectivityof these compounds from HSP90 inhibitors to kinase inhibitors. The factthat more than fourteen percent of the compounds showed a kinaseinhibition of greater than 50% at 10 mM clearly supports the hypothesisthat RAL is a good scaffold for kinase inhibition.

Example 25 HSP 90 Inhibition Tests for Exemplary Compounds of theLibrary

Radicicol (2-1), pochonin D (2-85) and pochonin A (2-122), along withsome closely related analogs such as monocillin II (2-103) or the diolanalog of pochonin A (2-125) were first evaluated for HSP90 affinity ina competition assay with geldanamycin using a previously describedmethod (V. Zhou et al., Anal. Biochem., 331, 349 (2004)). The resultsare shown in below.

As aforementioned, pochonin D (2-85) was found to be a good ligand forHSP90 with an IC₅₀ of 80 nM, as compared to 20 nM for radicicol. Thisdifference of activity is less than an order of magnitude, but withoutbeing bound by theory, it was rationalized by a molecular modellingstudy on pochonin D. The co-crystal structure of HSP90-radicicolreported by Pearl and co-workers showed a tightly bound water moleculemaking a hydrogen-bond bridge between the ortho-phenol, the ester, andAsp79 and a second water molecule making a bridge between thepara-phenol and Leu34 (S. M. Roe et al., J. Med. Chem., 42:260 (1999).Consistent with this hypothesis, compounds 2-112 and 2-120 withprotected phenols showed no affinity for HSP90. The importance of thechlorine atom is also evident from the comparison of pochonin D (2-85,80 nM) and monocillin II (2-103, >50 μM). The bulky chlorine not onlycomes within van der Waals contact of Phe124 but fills a hydrophobicpocket. Notably, pochonin A (2-122) was a good ligand (90 nM), whileanalog 2-111 was inactive. The comparison between radicicol, pochonin Dand pochonin A confirmed that the epoxide moiety is not essential forHSP90 inhibition and established that the γ,δ-conjugated olefin is not aprerequisite for potency. The ability of these natural products (2-1,2-85, 2-122) to antagonize ATP in the N-terminal domain of HSP90 and toinhibit ATPase activity may be related to their inhibition of HSV(Herpes Simplex Virus) helicase. Similarly, compounds 2-164, 2-166 and2-169 corresponding to bis-methylated analogs were evaluated for HSP90inhibition.

Example 26 Cytotoxicity Against HCC1954 and SK-BR-3 Tumor Cells forExemplary Compounds of the Library

Tumor cells were grown as adherent monolayers at 37° C. in a humidifiedatmosphere (5% CO₂, 95% air). The culture medium was RPMI 1640containing 2 mM L-glutamine (Ref BE12-702F, Cambrex, Verviers, Belgium)and supplemented with 10% fetal bovine serum (Ref DE14-801E, Cambrex).For experimental use, the adherent tumour cells were detached from theculture flask by a 5-minute treatment with trypsin-versene (Ref 02-007E,Cambrex), diluted in Hanks' medium without calcium or magnesium (RefBE10-543F, Cambrex) and neutralized by addition of complete culturemedium. Before use, cells were counted in a hemocytometer and theirviability were assessed by 0.25% trypan blue exclusion. Mycoplasmadetection was performed using the MycoAlert® Mycoplasma Detection Kit(Ref LT07-318, Cambrex) in accordance with the manufacturerinstructions. The MycoAlert® Assay is a selective biochemical test thatexploits the activity of mycoplasmal enzymes. The viable mycoplasma arelysed and the enzymes react with the MycoAlert® substrate catalyzing theconversion of ADP to ATP. By measuring the level of ATP in a sample bothbefore and after the addition of the MycoAlert® substrate a ratio can beobtained which is indicative of the presence or absence of mycoplasma.The mycoplasma test was assayed in duplicate from the culturesupernatants of the cell lines and compared to negative and positivecontrols (MycoAlert® Assay Control Set, Ref LT07-518, Cambrex) (InternalStandard Operating Procedure No TEC-007/002). Both HCC1954 and SK-BR-3tumour cells (5,000 cells per well) were plated in 96-well flat-bottommicrotitration plates (Ref 167008, Nunc, Dutscher, Brumath, France) andincubated at 37° C. for 24 hours before treatment in 190 μl of drug-freeculture medium supplemented with 10% FBS. Both HCC1954 and SK-BR-3tumour cell lines were incubated for 72 hours with 5 concentrations, in¼ dilution steps, of the test substances (ranging from 10⁻⁶ to 10⁻¹¹ M)to be tested as well as paclitaxel (ranging from 10⁻⁷ to 10⁻¹² M). Thecells (190 μl) will be incubated in a 200 μl final volume of culturemedium supplemented with 10% FBS containing test substances at 37° C.under 5% CO₂. One experiment is performed, each concentration beingissued from quadruplicate. Control cells are treated with correspondingvehicle alone. At the end of treatments, the cytotoxic activity isevaluated by a MTS assay. The in vitro cytotoxic activity of the testsubstances were revealed by a MTS assay using tetrazolium compound (MTS,3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) and an electron couplingreagent named PMS (phenazine methosulfate). Like MTT, MTS is bioreducedby cells into a formazan product that is directly soluble in culturemedium without processing, unlike MTT.

At the end of the cells treatment, 40 μl of a 0.22 μm filtered freshlycombined solution of MTS (20 ml at 2 mg/ml, Ref G1111A, Promega,Charbonnières, France) and PMS (1 ml at 0.92 mg/ml, Ref P9625, Sigma) inDulbecco's Phosphate Buffered Saline (DPBS, Ref 17-513F, Cambrex), wereadded in each well. Culture plates were incubated for 2 h at 37° C.Absorbency (OD) will be measured at 490 nm in each well usingVICTOR^(3TM) 1420 multilabeled counter (Wallac, PerkinElmer,Courtaboeuf, France).

TABLE 5 Cytotoxicity of selected pochonin analogs against HCC1954 andSK-BR-3 Tumor Cells Structure EC₅₀ HCC1954 EC₅₀ SK-BR-3 2a-1 

320 nM 125 nM E-isomer 2a-22

9.8 μM 3.8 μM 1:1 E:Z oxime mixture 2a-3 

3.0 μM 4.5 μM 1:1 E:Z oxime mixture 2a-4 

>10 μM 7.5 μM 2a-5 

9.5 μM 2.5 μM 2a-6 

5.2 μM >10 μM 2a-7 

>10 μM >10 μM 2a-8 

>10 μM >10 μM 2a-9 

890 nM 450 nM E isomer 2a-10

>10 μM >10 μM 2a-11

520 nM 310 nM 1:1 E:Z oxime mixture 2a-12

>10 μM 5.2 μM 2a-12

>10 μM >10 μM 1:1 E:Z oxime mixture 2a-13

220 nM 120 nM E isomer 2a-14

1.2 μM 2.8 μM 2a-15

>10 μM >10 μM 1:1 E:Z oxime mixture 2a-16

450 nM 550 nM 1:1 E:Z oxime mixture 2a-17

8.5 μM 3.5 μM 1:1 E:Z oxime mixture 2a-18

>10 μM >10 μM 1:1 E:Z oxime mixture

Experimental procedures and characterization data for exemplarycompounds follows. The compounds and data described below are understoodto be non-limiting.

General procedure for oxidation using solvent system A: A solution ofaldehyde 2-94 (1.0 equiv.) in H₂O/THF/DMSO (20:10:1, 0.03 M) wassequentially treated at 0° C. with sulfamic acid (3.5 equiv.) and asolution of sodium chlorite (3.25 equiv.) in H₂O. After 0.5-1 h stirringat this temperature, the reaction mixture was diluted with Et₂O, washedwith saturated NH₄Cl_(aq.) and dried over MgSO₄. Concentration underreduced pressure afforded the corresponding acid 2-95 which was usedwithout any further purification in the next step.

2-95c: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.): δ=6.48 (d, J=2.2 Hz, 1H),6.45 (d, J=2.2 Hz, 1H), 3.84 (s, 3H), 3.83 (s, 3H), 2.33 (s, 3H).

2-94j: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.): δ=7.58 (d, J=7.0 Hz, 4H),7.47-7.35 (m, 10H), 7.31-7.21 (m, 6H), 6.31 (s, 1H), 5.89 (s, 1H), 2.35(s, 3H), 1.05 (s, 9H), 0.93 (s, 9H).

General procedure for oxidation using solvent system B: A solution ofaldehyde 2-94 (1.0 equiv.) in H₂O/THF (20:10, 0.03 M) was sequentiallytreated at 0° C. with sulfamic acid (3.5 equiv.) and a solution ofsodium chlorite (3.25 equiv.) in H₂O. After 12 h stirring at roomtemperature, the reaction mixture was diluted with Et₂O, washed withsaturated NH₄Cl_(aq.) and dried over MgSO₄. Concentration under reducedpressure afforded the corresponding acid 2-95 which was used without anyfurther purification in the next step. For 2-95b, 2.0 equiv. of sulfamicacid were necessary to avoid over-chlorination. For 2-95i and 2-95k, thereaction was complete after 30 min at 0° C.

2-95b: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.): δ=6.48 (s, 1H), 2.70 (s, 3H).

2-95d: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.): δ=6.77 (s, 1H), 3.97 (s, 3H),3.89 (s, 3H), 2.35 (s, 3H).

2-95h: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.): δ=6.71 (s, 1H), 5.43 (s, 2H),3.85 (t, J=8.2 Hz, 2H), 2.71 (s, 3H), 1.00 (t, J=8.2 Hz, 2H), 0.04 (s,9H).

2-95i: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.): δ=7.82-7.80 (m, 4H),7.55-7.48 (m, 6H), 6.02 (s, 1H), 2.74 (s, 3H), 1.17 (s, 9H).

2-95k: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.): δ=7.48-7.34 (m, 12H),7.23-7.16 (m, 8H), 5.93 (s, 1H), 2.54 (s, 3H), 1.05 (s, 9H), 1.02 (s,9H).

General procedure for oxidation using solvent system C: To a solution ofaldehyde 2-94 (1.0 equiv.) in DMSO (0.4 M) at 0° C., were added slowlyin a sequential fashion, NaH₂PO₄.H₂O (5.0 equiv.) dissolved in H₂O (3 M)and NaClO₂ (5.0 equiv.) dissolved in H₂O (3 M). After stirring for 12 h,the reaction was diluted with Et₂O, washed with saturated NH₄Cl_(aq.)and dried over MgSO₄. Concentration under reduced pressure resulted intothe corresponding acid 2-95 used without further purification in thenext step.

2-95e: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.): δ=6.76 (d, J=1.9 Hz, 1H),6.60 (d, J=1.6 Hz, 1H), 5.25 (s, 4H), 3.75-3.69 (m, 4H), 2.31 (s, 3H),1.18 (t, J=7.0 Hz, 6H).

2-95g: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.): δ=12.13 (bs, 1H), 6.46 (s,2H), 5.30 (s, 2H), 3.73 (q, J=7.0 Hz, 4H), 2.58 (s, 3H), 1.19 (t, J=7.0Hz, 3H).

1-Chloro-2,4-bis-ethoxymethoxy-7-methyl-7,8,11,12-tetrahydro-16H-6-oxa-benzocyclotetradecene-5,15-dione(2-112): A 2 mM solution of compound 2-111 (200 mg, 0.38 mmol) inanhydrous toluene (190 mL) was treated with 10% mol of catalyst Grubbs'II (30 mg, 0.038 mmol) and heated at 80° C. overnight. The reactionmixture was then passed through a pad of silica, which was washed withCH₂Cl₂. The combined filtrates were concentrated under reduced pressure.Purification by flash chromatography (silica gel, 0-25% EtOAc/hexanegradient) afforded pure macrocycle 2-112 (167 mg, 94%). ¹H NMR (400 MHz,C₆D₆, 25° C.): δ=7.27 (s, 1H), 6.85 (dt, J=15.2, 7.6 Hz, 1H), 6.15 (d,J=15.8 Hz, 1H), 5.16-4.94 (m, 7H), 4.41 (d, J=17.0 Hz, 1H), 4.13 (d,J=17.0 Hz, 1H), 3.61-3.45 (m, 4H), 2.18-2.07 (m, 2H), 1.86-1.62 (m, 4H),1.38 (d, J=5.8 Hz, 3H), 1.11 (t, J=7.0 Hz, 3H), 1.04 (t, J=7.0 Hz, 3H);¹³C NMR (100 MHz, C₆D₆, 25° C.): δ=193.5, 166.4, 155.0, 154.1, 146.0,133.7, 131.5, 128.8, 127.7, 121.2, 118.1, 102.9, 93.7, 93.6, 71.6, 64.4,64.4, 45.0, 39.2, 30.7, 30.4, 19.3, 14.9, 14.8; I.R. (film):ν_(max)=2917, 1720, 1690, 1622, 1591, 1320, 1255, 1120, 1037 cm⁻¹; HRMS(ESI-TOF): m/z: calculated for C₂₄H₃₁O₇ClNa: 489.1651, found 489.1737[M+Na⁺]. (−)-(2R): [α]²⁵ _(D)=−24.0 (c 0.59, CHCl₃).

Pochonin D (2-85): Compound 2-112 (50 mg, 0.1 mmol) was stirred for 2 hin a 5:1 mixture of CH₂Cl₂/TFA (3 mL). Concentration under reducedpressure, followed by flash chromatography (silica gel, 0-33%EtOAc/hexane gradient) afforded synthetic pochonin D 2-85 (25 mg, 72%).Synthetic pochonin D was found to have identical ¹H NMR as naturalpochonin D. ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=12.42 (s, 1H), 6.89 (s,1H), 6.67-6.62 (m, 1H), 5.82 (d, J=15.6 Hz, 1H), 5.17-5.12 (m, 1H),5.00-4.92 (m, 1H), 4.76-4.69 (m, 1H), 4.28 (d, J=17.2 Hz, 1H), 4.18 (d,J=17.7 Hz, 1H), 2.54-2.47 (m, 1H), 1.93-1.77 (m, 5H), 0.98 (d, J=6.4 Hz,3H); ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=11.73 (s, 1H), 6.76-6.69 (m+s,2H), 6.19 (s, 1H), 5.82 (d, J=15.2 Hz, 1H), 5.46-5.40 (m, 1H), 5.31-5.15(m, 2H), 4.37 (d, J=17.6 Hz, 1H), 4.09 (d, J=17.0 Hz, 1H), 2.68-2.61 (m,1H), 2.39-2.03 (m, 5H), 1.34 (d, J=7.0 Hz, 3H); ¹H NMR (400 MHz, CD₃OD,25° C.): δ=6.74 (dt, J=15.5, 7.6 Hz, 1H), 6.51 (s, 1H), 5.83 (d, J=15.5Hz, 1H), 5.36-5.22 (m, 3H), 4.25 (d, J=17.7 Hz, 1H), 4.13 (d, J=17.7 Hz,1H), 2.54-2.47 (ddd, J=14.5, 8.0, 4.0 Hz, 1H), 2.31-2.15 (m, 5H), 1.31(d, J=6.6 Hz, 3H); ¹³C NMR (100 MHz, C₆D₆, 25° C.): δ=194.2, 169.9,164.3, 157.3, 146.1, 137.1, 131.9, 128.1, 126.2, 115.5, 107.5, 103.6,72.4, 45.0, 36.4, 31.0, 30.8, 17.2; I.R. (KBr): ν_(max)=2936, 1654,1603, 1347, 1313, 1239 cm⁻¹; HRMS (ESI-TOF): m/z: calculated forC₁₈H₁₉O₅ClNa: 373.0813, found 373.0903 [M+Na⁺]. (+)-(2R): [α]²⁵_(D)=+11.1 (c 0.72, CHCl₃).

Hepta-2,6-dienoic acid methoxy-methyl-amide (2-114): To a solution of2-chloro-N-methoxy-N-methylacetamide (6.0 g, 48.8 mmol) in dry DMF (20mL) at 23° C. was added 3-mercaptophenol (4.44 mL, 48.8 mmol) and K₂CO₃(6.7 g, 48.8 mmol). The resulting suspension was stirred at 23° C.overnight. After this period of time, Merrifield resin (24 g, <2mmol.g⁻¹, <48.8 mmol) was added to the mixture followed by K₂CO₃ (11.4g, 83.0 mmol) as well as TBAI (catalytic amount), and the suspension washeated up to 50° C. After 12 hours at this temperature, the resin wasfiltered and washed several times: HCl_(aq.) (50 mL), MeOH (50 mL),CH₂Cl₂ (50 mL) and Et₂O (50 mL). The resin was dried under reducedpressure to constant mass of 29.2 g. The final mass gain (5.2 g, 27.3mmol) indicated an estimate loading of 0.81 mmol.g⁻¹. Resin 2-49 (10 g,0.81 mmol.g⁻¹) was suspended in a 1:1 mixture of HFIP/CH₂Cl₂ (50 mL). Tothis suspension, H₂O₂ (3 mL, 16.0 mmol) was added at 23° C. and theresulting mixture was shaken for 12 h. Resin 2-113 was then filtered,washed using MeOH (50 mL), CH₂Cl₂ (50 mL) and Et₂O (50 mL) and driedunder reduced pressure to constant mass before subsequent use. Resin2-113 (4.0 g, <0.81 mmol.g⁻¹) was suspended in DMSO (40 mL) followed bythe addition of tBuOK (336 mg, 3.0 mmol). After shaking the reaction for1 h at room temperature, 5-iodo-1-pentene (588 mg, 3.0 mmol) was addedto the suspension and the mixture was shaken for 3 h. The resin wasfiltered, washed and dried as before. Then, it was suspended in tolueneand heated at 80° C. After 8 h at this temperature, the resin wasfiltered and washed several times with more toluene. The combinedtoluene solutions were evaporated giving pure compound 2-114 as acolourless oil (321 mg, 77%) of 95% purity judged by NMR. ¹H NMR (400MHz, CDCl₃, 25° C.): δ=6.94 (dt, J=15.7, 6.7 Hz, 1H), 6.39 (d, J=15.2Hz, 1H), 5.84-5.74 (m, 1H), 5.02 (dd, J=17.4, 1.7 Hz, 1H), 4.97 (d,J=10.1 Hz, 1H), 3.67 (s, 3H), 3.21 (s, 3H), 2.34-2.29 (m, 2H), 2.23-2.18(m, 2H); ¹³C NMR (100 MHz, CDCl₃, 25° C.): δ=166.8, 146.7, 137.3, 119.1,115.3, 61.6, 32.3, 31.7, (one carbon is not detected); I.R. (film):ν_(max)=2934, 1681, 1638, 1378, 1179 cm⁻¹.

2,4-Bis-ethoxymethoxy-7-methyl-7,8,11,12-tetrahydro-16H-6-oxa-benzocyclotetradecene-5,15-dione(2-120). A 2 mM solution of crude 2-119 (1.5 mmol) in anhydrous toluene(750 mL) was treated with 10% mol of catalyst Grubbs' II (139 mg, 0.15mmol), and heated at 80° C. overnight. The crude reaction mixture wasthen passed through a pad of silica, which was washed with CH₂Cl₂. Thecombined filtrates were concentrated under reduced pressure.Purification by flash chromatography (silica gel, 0-25%EtOAc/cyclohexane gradient) afforded pure 2-120 (260 mg, 40% over twosteps). ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=7.08 (d, J=2.2 Hz, 1H), 7.01(d, J=2.2 Hz, 1H), 6.93-6.86 (m, 1H), 6.17 (d, J=16.1 Hz, 1H), 5.34-4.90(m, 7H), 4.41 (d, J=14.5 Hz, 1H), 3.75 (d, J=14.5 Hz, 1H), 3.59-3.45 (m,4H), 2.27-2.12 (m, 2H), 1.95-1.61 (m, 4H), 1.45 (d, J=6.2 Hz, 3H), 1.07(t, J=7.0 Hz, 3H), 1.07 (t, J=7.0 Hz, 3H); ¹³C NMR (100 MHz, C₆D₆, 25°C.): δ=196.0, 167.2, 159.5, 156.6, 147.6, 135.8, 131.5, 130.2, 128.5,119.1, 109.7, 102.3, 93.3, 92.9, 71.0, 64.2, 64.0, 44.5, 39.6, 30.9,30.2, 20.1, 14.8, 14.8; I.R. (film): ν_(max)=2976, 1717, 1602, 1438,1284, 1155, 1110, 1036, 1018 cm⁻¹; HRMS (ESI-TOF): nm/z: calculated forC₂₄H₃₂O₇Na: 455.2040, found 455.2135 [M+Na⁺]. (−)-(2R): [α]²⁵ _(D)=−50.3(c 1.00, CHCl₃).

Pochonin D (2-85) using polymer-bound reagents: PS-TsOH (300 mg, 3.2mmol.g⁻¹) was added to a solution of compound 2-112 (50 mg, 0.1 mmol) inMeOH (3 mL) of and the suspension was shaken at 40° C. for 4 h. Thereaction mixture was then filtered and the methanolic solutionconcentrated under reduced pressure. Purification by flashchromatography (silica gel, 0-20% EtOAc/cyclohexane gradient) affordedsynthetic pochonin D 2-85 (32 mg, 90%).

Monocillin II (2-103) using polymer-bound reagents: PS-TsOH (145 mg, 3.2mmol.g⁻¹) was added to a solution of compound 2-120 (20 mg, 0.05 mmol)in MeOH (1.5 mL) and the suspension was shaken at 40° C. for 4 h. Thereaction mixture was then filtered and the methanolic solutionconcentrated under reduced pressure. Purification by flashchromatography (silica gel, 0-20% EtOAc/cyclohexane gradient) affordedsynthetic monocillin II 2-103 (14 mg, 92%). ¹H NMR (400 MHz, C₆D₆, 25°C.): δ=12.49 (s, 1H), 6.70-6.62 (m, 1H), 6.49 (d, J=2.9 Hz, 1H), 6.09(d, J=2.4 Hz, 1H), 5.86 (d, J=15.2 Hz, 1H), 5.08-4.95 (m, 2H), 4.82-4.75(m, 1H), 4.14 (d, J=16.8 Hz, 1H), 3.71 (d, J=17.0 Hz, 1H), 2.64-2.57 (m,1H), 1.83-1.76 (m, 3H), 1.74-1.66 (m, 2H), 0.97 (d, J=6.4 Hz, 3H); ¹³CNMR (100 MHz, C₆D₆, 25° C.): δ=195.7, 170.3, 166.5, 161.3, 146.1, 140.5,131.7, 129.9, 126.5, 112.4, 102.8, 72.2, 49.1, 36.6, 31.0, 30.6, 17.5,(one carbon is not detected); I.R. (KBr): ν_(max)=2936, 1654, 1603,1347, 1313, 1239 cm⁻¹; HRMS (ESI-TOF): m/z: calculated for C₁₈H₂₁O₅requires 317.3980, found 317.3978 [M+H⁺]. (+)-(2R): [α]²⁵ _(D)=+40.6 (c0.18, CHCl₃).

Macrocycle 2-122: To a solution of compound 2-112 (50 mg, 0.11 mmol) inCH₃CN (5 mL) at 0° C. was added freshly made DMDO (275 μL, 0.11 mmol,0.04 M in acetone) and the mixture was stirred for 1.5 h. Afterevaporation of the solvents under reduced pressure, purification byflash chromatography (silica gel, 0-70% Et₂O/hexane gradient) affordedcompound 2-122 (41 mg, 79%) as a 1:1 mixture of two diastereoisomers. ¹HNMR (400 MHz, CDCl₃, 25° C.): δ=7.12 (s, 1H), 7.11 (s, 1H), 6.90-6.76(m, 2H), 6.11 (d, J=15.6 Hz, 1H), 6.05 (d, J=15.8 Hz, 1H), 5.37-5.27 (m,6H), 5.26-5.21 (m, 4H), 4.14 (d, J=16.9 Hz, 1H), 4.12 (d, J=17.4 Hz,1H), 4.04 (d, J=17.2 Hz, 1H), 3.82-3.70 (m, 9H), 2.81-2.78 (m, 1H),2.74-2.72 (m, 1H), 2.67-2.62 (m, 2H), 2.38-2.11 (m, 8H), 2.05-2.03 (m,1H), 2.03-2.00 (m, 1H), 1.74-1.60 (m, 2H), 1.41 (d, J=7.2 Hz, 3H), 1.39(d, J=6.2 Hz, 3H), 1.27-1.22 (m, 12H); ¹³C NMR (100 MHz, CDCl₃, 25° C.):δ=195.1 (×2), 166.7, 166.3, 154.9, 154.8, 154.0, 153.5, 147.8, 147.4,132.9, 132.4, 129.1, 129.0, 119.7 (×2), 118.0, 117.9, 102.9, 102.8, 93.9(×2), 93.6 (×2), 71.1, 70.4, 64.8 (×2), 64.6 (×2), 58.4, 57.6, 56.9,55.1, 43.4 (×2), 39.0, 37.9, 29.9, 29.7, 27.9 (×2), 18.4, 18.0, 15.0(×4); HRMS (ESI-TOF): m/z: calculated for C₂₄H₃₂ClO₈: 483.1780, found483.1814 [M+H⁺].

Macrocycles 2-124 and 2-125: PS-TsOH (264 mg, 3.2 mmol.g⁻¹) was added toa solution of compound 2-123 (41 mg, 85 mmol) in MeOH (3 mL) and thesuspension was shaken at 40° C. until consumption of all startingmaterial (˜1 h). The reaction mixture was filtered and the methanolicsolution concentrated under reduced pressure. L.C./M.S. analysis of thecrude mixture showed clearly 2 peaks corresponding to methanol additionon the conjugated olefin (2-124) and opening of the epoxide as a diol(2-125).

2-125: ¹H NMR (400 MHz, CD₃OD, 25° C.): δ=7.19 (m, 1H), 6.89-6.81 (m,1H), 6.52 (s, 1H), 6.47 (s, 1H), 6.20 (d, J=16.1 Hz, 1H), 6.04 (d,J=15.6 Hz, 1H), 5.54-5.49 (m, 1H), 5.43-5.36 (m, 1H), 4.50 (d, J=17.7Hz, 1H), 4.46 (d, J=17.7 Hz, 1H), 4.39 (d, J=17.2 Hz, 1H), 4.07 (d,J=17.2 Hz, 1H), 3.80-3.64 (m, 2H), 3.51-3.46 (m, 2H), 2.62-2.58 (m, 1H),2.39-2.30 (m, 2H), 2.27-2.18 (m, 1H), 2.08-2.00 (m, 2H), 2.00-1.85 (m,4H), 1.44 (d, J=6.4 Hz, 6H); HRMS (ESI-TOF): m/z: calculated forC₁₈H₂₂ClO₇: 385.1054, found 385.0944 [M+H⁺].

2-124: compound 2-124 which was characterized as the MeOH-addition onthe α,β-conjugated system based on the loose of olefinic protons in theNMR (a detailed assignment is not possible as product 2-124 represents amixture of 4 compounds); HRMS (ESI-TOF): m/z: calculated for C₁₉H₂₄ClO₇:399.1211, found 399.1030 [M+H⁺].

Macrocycle 2-128: A 2 mM solution of compound 2-127 (140 mg, 0.23 mmol)in anhydrous toluene (115 mL) was treated with 10% mol of catalystGrubbs' II (18.4 mg, 0.023 mmol) and heated up to 80° C. for 12 h. Thereaction mixture was then filtered through a pad of silica, which waswashed with CH₂Cl₂. The combined filtrates were concentrated underreduced pressure. Purification by flash chromatography (silica gel,0-25% EtOAc/hexane gradient) afforded macrocycle 2-128 (116 mg, 87%). ¹HNMR (400 MHz, CDCl₃, 25° C.): 8=6.71 (dt, J=15.3, 7.3 Hz, 1H), 6.45 (s,1H), 5.81 (d, J=15.3 Hz, 1H), 5.25 (s, 2H), 5.04-5.03 (m, 1H), 3.89 (d,J=17.4 Hz, 1H), 3.57 (d, J=17.4 Hz, 1H), 2.31-2.04 (m, 6H), 1.35 (d,J=6.4 Hz, 3H), 1.03 (s, 9H), 0.99 (s, 9H), 0.28-0.24 (m, 12H); ¹³C NMR(100 MHz, CDCl₃, 25° C.): δ=195.8, 166.8, 152.9, 151.7, 146.5, 132.7,131.9, 128.6, 126.8, 122.8, 119.7, 110.7, 71.9, 45.6, 38.5, 30.9, 25.7(×4), 25.6 (×4), 18.7, 18.3, −4.1 (×2), −4.4 (×2); HRMS (ESI-TOF): m/z:calculated for C₃₀H₄₇ClO₅Si₂Na: 601.2543, found 601.2568 [M+Na⁺].

Macrocycle 2-129: An aqueous Na₂.EDTA solution (700 μL, 4×10⁻⁴M) wasadded to a solution of compound 2-128 (80 mg, 0.14 mmol) in a 2:1mixture of dimethoxymethane/acetonitrile (2.1 mL). The resulting mixturewas cooled to 0° C. and treated with trifluoroacetone (150 μL) added viaa precooled syringe. A mixture of sodium bicarbonate (88 mg, 1.05 mmol)and Oxone (430 mg, 0.70 mmol) was added in portions over a period of ˜1h to this homogeneous solution. The reaction was followed by TLC andfound to be complete in 2 h. The reaction mixture was then poured intowater (10 mL), extracted with CH₂Cl₂ (20 mL) and dried over Na₂SO₄.Removal of the solvent under reduced pressure afforded pure compound2-129 (66 mg, 93%) as a mixture of 2 diastereoisomers in a 3:1 ratio. ¹HNMR (400 MHz, CDCl₃, 25° C.): δ=6.90-6.77 (m, 1.33H), 6.45 (s, 1.33H),6.06 (d, J=15.8 Hz, 1.33H), 5.31-5.29 (m, 1H), 5.29-5.21 (m, 0.33H),4.03 (d, J=18.1 Hz, 1.33H), 3.63 (d, J=17.6 Hz, 1H), 2.82-2.80 (m,1.33H), 2.74-2.71 (m, 1.33H), 2.62-2.60 (m, 1.33H), 2.41-2.11 (m, 4.6H),2.02-1.95 (m, 0.33H), 1.80-1.78 (m; 1H), 1.78-1.68 (m, 0.7H), 1.41 (d,J=6.4 Hz, 3.9H), 1.05 (s, 12H), 0.97 (s, 12H), 0.26 (s, 16H); ¹³C NMR(100 MHz, CDCl₃, 25° C.): δ=195.4, 195.1, 167.0, 166.2, 153.1 (×2),152.1, 151.5, 147.8, 146.9, 132.9, 132.2, 129.0, 128.3, 122.2, 121.4,120.0, 119.7, 110.6, 110.3, 71.3, 70.1, 58.2, 57.9, 56.6, 55.2, 44.7,43.9, 38.7, 37.6, 30.1, 29.4, 28.3, 27.5, 25.7 (×4), 25.6 (×4), 25.5(×4), 25.4 (×4), 20.8, 17.8, −3.9, −4.0, −4.3 (×3), −4.4 (×3); HRMS(ESI-TOF): m/z: calculated for C₃₀H₄₈O₆ClSi₂: 595.2672, found 595.2698[M+H⁺].

Pochonin A (2-122): TBAF (244 μL, 1M solution in hexane, 0.24 mmol) wasadded to a solution of compound 2-129 (66 mg, 0.11 mmol) in THF (2 mL)and the mixture was stirred at room temperature for 20 min. The reactionwas then quenched with saturated NH₄Cl_(aq.) (8 mL), extracted severaltimes with EtOAc (10 mL) and dried over Na₂SO₄. Concentration underreduced pressure followed by purification by flash chromatography(silica gel, 0-70% Et₂O/hexane) afforded two different diastereoisomerspochonin A (2-122) and its diastereoisomer 2-122b as a 3:1 mixture (80%yield) The isomers were separated by preparative TLC with a 3:1 mixtureof Et₂O/hexane. ¹H NMR (400 MHz, [D₆]DMSO, 25° C.): δ=10.81 (s, 1H),10.74 (s, 1H), 6.97-6.89 (m, 1H), 6.53 (s, 1H), 6.08 (d, J=15.8 Hz, 1H),5.15-5.13 (m, 1H), 4.19 (d, J=17.5 Hz, 1H), 4.09 (d, J=17.5 Hz, 1H),2.81 (s, 1H), 2.60 (m, 1H), 2.44-2.40 (m, 2H), 2.30-2.22 (m, 2H),1.80-1.78 (m, 2H), 1.32 (d, J=6.4 Hz, 3H); ¹H NMR (400 MHz, CDCl₃, 25°C.): δ=11.85 (s, 1H), 6.94-6.87 (m, 1H), 6.70 (s, 1H), 6.14 (s, 1H),6.12 (d, J=16.4 Hz, 1H), 5.32-5.31 (m, 1H), 4.53 (d, J=18.1 Hz, 1H),4.27 (d, J=18.1 Hz, 1H), 2.77 (s, 1H), 2.58-2.56 (m, 2H), 2.47-2.43 (m,1H), 2.35-2.28 (m, 1H), 2.11-2.07 (m, 1H), 1.93-1.86 (m, 1H), 1.51 (d,J=6.4 Hz, 3H), 0.94-0.90 (m, 1H); ¹³C NMR (100 MHz, CDCl₃, 25° C.):δ=195.0, 170.0, 164.1, 156.4, 147.5, 135.7, 129.9, 115.0, 107.3, 103.8,72.2, 57.0, 55.5, 45.1, 36.3, 30.9, 29.1, 17.9; HRMS (ESI-TOF): m/z:calculated for C₁₈H₁₉ClO₆Na: 389.0762, found 389.0724 [M+Na⁺].(−)-(2R,4R, SR): [α]²⁵ _(D)=−7.0 (c 0.11, CHCl₃).

Compound 2-122b: ¹H NMR (400 MHz, [D₆]DMSO, 25° C.): δ=10.74 (s, 1H),10.39 (s, 1H), 6.94-6.89 (m, 1H), 6.52 (s, 1H), 6.02 (d, J=16.4 Hz, 1H),5.18 (m, 1H), 4.32 (d, J=17.5 Hz, 1H), 3.96 (d, J=17.5 Hz, 1H), 2.82 (s,1H), 2.68 (s, 1H), 2.34-2.26 (m, 3H), 1.86-1.83 (m, 1H), 1.70-1.63 (m,1H), 1.22 (d, J=5.8 Hz, 3H), 1H masked by the solvent peak; ¹H NMR (400MHz, CDCl₃, 25° C.): δ=11.37 (s, 1H), 6.90-6.83 (m, 1H), 6.67 (s, 1H),6.24 (d, J=16.4 Hz, 1H), 6.08 (s, 1H), 5.39-5.37 (m, 1H), 4.52-4.36 (m,2H), 2.72-2.62 (m, 2H), 2.56-2.52 (m, 1H), 2.45-2.40 (m, 1H), 2.40-2.37(m, 1H), 2.08-2.04 (m, 1H), 1.91-1.86 (m, 1H), 1.35 (d, J=6.4 Hz, 3H),1H masked by the solvent peak; HRMS (ESI-TOF): m/z: calculated forC₁₈H₁₉ClO₆Na: 389.0762; found 389.0796 [M+Na⁺]. (+)-(2R,4S,5S): [α]²⁵_(D)=+13.8 (c 0.13, CHCl₃).

Macrocycle 2-132: A 2 mM solution of compound 2-131 (166 mg, 0.26 mmol)in anhydrous toluene (130 mL) was treated with 10% mol of Grubbs' II(20.8 mg, 0.026 mmol) and heated up to 80° C. for 12 h. The reactionmixture was then filtered through a pad of silica, which was washed withCH₂Cl₂. The combined filtrates were concentrated under reduced pressure.Purification by flash chromatography (silica gel, 0-25% EtOAc/hexanegradient) afforded macrocycle 2-132 (136 mg, 87%). ¹H NMR (400 MHz,CDCl₃, 25° C.): δ=7.08 (s, 1H), 6.77-6.71 (m, 1H), 5.89 (d, J=15.2 Hz,1H), 5.35-5.24 (m, 6H), 5.09-5.05 (m, 1H), 4.02 (d, J=17.0 Hz, 1H),3.85-3.78 (m, 5H), 2.37-2.08 (m, 6H), 1.39 (d, J=5.8 Hz, 3H), 1.02-0.97(m, 4H), 0.04 (s, 18H); ¹³C NMR (100 MHz, CDCl₃, 25° C.): δ=195.7,166.8, 154.6, 153.9, 147.0, 132.8, 131.7, 128.6, 127.5, 120.6, 117.8,102.7, 93.7, 93.3, 71.9, 66.9, 66.6, 44.7, 39.2, 30.8 (×2), 19.5, 18.0,17.9, −1.4 (×6); HRMS (ESI-TOF): m/z: calculated for C₃₀H₄₇O₇ClSi₂H₂O:628.2649, found 628.2870 [M+H₂O]. (−)-(2R): [α]²⁵ _(D)=−16.3 (c 0.85,CHCl₃).

Macrocycle 2-133: An aqueous Na₂.EDTA solution (350 μL, 4×10⁻⁴M) wasadded to a solution of compound 2-132 (40 mg, 65 μmol) in a 2:1 mixtureof dimethoxymethane/acetonitrile (1.1 mL). The resulting solution wascooled to 0° C. and trifluoroacetone (75 μL) was added via a precooledsyringe. A mixture of sodium bicarbonate (44 mg, 0.5 mmol) and Oxone(215 mg, 0.35 mmol) was then added in portions over a period of ˜1 h tothis homogeneous solution. The reaction was followed by TLC and found tobe complete in 2 h. The reaction mixture was then poured into water (5mL), extracted with CH₂Cl₂ (10 mL) and dried over Na₂SO₄. Removal of thesolvent under reduced pressure afforded pure compound 2-133 (34 mg, 82%)as a 1:1 mixture of two diastereoisomers. ¹H NMR (400 MHz, CDCl₃, 25°C.): δ=7.11 (s, 1H), 7.09 (s, 1H), 6.91-6.76 (m, 2H), 6.12 (d, J=15.8Hz, 1H), 6.06 (d, J=15.8 Hz, 1H), 5.36-5.31 (m, 6H), 5.23-5.22 (m, 4H),4.15 (d, J=16.9 Hz, 1H), 4.13 (d, J=16.0 Hz, 1H), 4.05 (d, J=17.5 Hz,1H), 3.85-3.75 (m, 9H), 2.81-2.79 (m, 1H), 2.75-2.73 (m, 1H), 2.68-2.62(m, 2H), 2.42-2.29 (m, 10H), 1.72-1.60 (m, 2H), 1.41 (d, J=7.6 Hz, 3H),1.39 (d, J=6.4 Hz, 3H), 1.02-0.96 (m, 8H), 0.03 (s, 36H); ¹³C NMR (100MHz, CDCl₃, 25° C.): δ=195.1 (×2), 166.8, 166.3, 154.9 (×2), 154.2,153.7, 147.8, 147.4, 132.9, 132.4, 129.1, 129.0, 119.8, 119.6, 117.9,117.8, 102.8, 102.7, 93.7 (×2), 93.4 (×2), 71.1, 70.3, 66.9, 66.8, 66.6,66.5, 58.4, 57.6, 56.9, 55.1, 43.5 (×2), 38.9, 37.9, 29.9, 29.7, 27.9(×2), 20.7 (×2), 18.4, 18.0, 17.9 (×2), −1.40 (×12); HRMS (ESI-TOF):m/z: calculated for C₃₀H₄₈O₈ClSi₂: 627.2571, found 627.2551 [M+H⁺].

Pochonin A (2-122) from macrocycle 2-133: A solution of compound 2-133(21 mg, 33 μmol) in CH₂Cl₂ (2.5 mL) was treated at room temperature withMgBr₂.Et₂O (69 mg, 0.27 mmol). The reaction was followed by L.C./M.S.until bromohydrine started appearing (˜1 h). The reaction was thendiluted with EtOAc (5 mL), washed with saturated NH₄Cl_(aq.) (5 mL) anddried over MgSO₄. After concentration under reduced pressure,purification by flash chromatography (silica gel, 0-70% Et₂O/hexanegradient) afforded pochonin A (2-122) (8.6 mg, 70%) as a 1:1 mixture ofdiastereoisomers.

General procedure for the synthesis of compounds 2-110a-g and 2-117a-g:A solution of acid 2-95a or 2-95b (1.0 equiv.), homoallylic alcohol(R)-120a-g or (S)-120a-g (1.0 equiv.) and tris-(3-chlorophenyl)phosphine(2.0 equiv.) in anhydrous toluene (0.05 M) was treated at roomtemperature with PS-DEAD (2.5 equiv., 1.3 mmol.g⁻¹). After stirring for30 min, the reaction mixture was filtered on silica and washed withhexane/EtOAc (10:1, 100 mL) and hexane/EtOAc (3:1, 100 mL). The 3:1mixture was concentrated under reduced pressure to yield compound2-115a-g or 2-116a-g (60-80%). Without further purification, compound2-115a-g or 2-116a-g (1.0 equiv.) and TBAI (catalytic amount) weredissolved in DMF (0.15 M) and treated with diisopropylethylamine (4.0equiv.) and chloromethylethyl ether (4.0 equiv.). After stirringovernight at 80° C., the reaction mixture was diluted with EtOAc andwashed several times with a saturated NH₄Cl_(aq.) solution. The organicphase was dried over MgSO₄ and concentrated under reduced pressure toyield compounds 2-110a-g and 2-117a-g (80-90%).

General procedure for the synthesis of compounds 2-119a-g and 2-140a-g:A solution of compound 2-110a-g or 2-117a-g (1.0 equiv.) in anhydrousTHF (0.2 M) cooled at −78° C. was treated with freshly prepared LDA (2.0equiv.). Immediately after, the α,β-unsaturated Weinreb amide 2-114 wasadded to the cooled solution (1.0 equiv.). The resulting mixture wasthen stirred for 10 min at −78° C. and quenched by addition of Amberliteresin (20 equiv.). Upon warming up to room temperature, the reaction wasfiltered on a pad of silica and washed with EtOAc. Concentration underreduced pressure afforded the desired compound 2-118a-g or 2-119a-g.This compound was used directly in the metathesis reaction without anyfurther purification. When X═H, 20% of the corresponding 1,4-additioncompound was observed and a fraction of the mixture was purified forcharacterization of compounds 2-119a-g and 2-140a-g (silica gel, 0-20%EtOAc/hexane gradient).

General procedure for the metathesis reaction: A solution of crude2-118a-g or 2-119a-g (or mixture 2-119a-g or 2-140a-g when X═H) inanhydrous toluene (2 mM) was treated with Grubbs' II (0.10 equiv.) andheated at 80° C. for 12 h. The reaction was cooled down to roomtemperature and the mixture was filtered through a pad of silica gel,washed with CH₂Cl₂ followed by a mixture EtOAc/cyclohexane 1:1, andconcentrated under reduced pressure. Purification by flashchromatography (silica gel, 0-25% EtOAc/cyclohexane gradient) affordedcompound 2-112a-g or 2-120a-g (and 2-121a-g) (38-70% over two steps).

Compound (S)-2-112: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=7.10 (s, 1H),6.75-6.71 (m, 1H), 5.88 (d, J=15.8 Hz, 1H), 5.32 (s, 2H), 5.27-5.20 (m,2H), 5.25 (s, 2H), 5.08-5.04 (m, 1H), 4.01 (d, J=17.0 Hz, 1H), 3.82-3.73(m, 5H), 2.36-2.32 (m, 2H), 2.26-2.20 (m, 3H), 2.12-2.05 (m, 2H), 1.38(d, J=5.8 Hz, 3H), 1.25 (t, J=7.0 Hz, 3H), 1.24 (t, J=7.0 Hz, 3H); ¹³CNMR (100 MHz, CDCl₃, 25° C.): δ=195.5, 166.7, 154.6, 153.8, 147.1,132.8, 131.6, 128.6, 127.5, 120.7, 117.8, 103.0, 94.0, 93.7, 72.0, 64.8,64.6, 44.7, 39.1, 30.8, 19.4, 15.0, 2C missing; HRMS (ESI-TOF): m/z:calculated for C₂₄H₃₁O₇ClNa: 489.1551, found 489.1651 [M+Na⁺]. (+)-(2S):[α]²⁵ _(D)=+25.0 (c 1.00, CHCl₃).

Compound (R)-2-112a: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=7.49-7.47 (m,2H), 7.40-7.29 (m, 3H), 7.10 (s, 1H), 6.84-6.77 (m, 1H), 5.98 (d, J=15.2Hz, 1H), 5.78 (d, J=8.8 Hz, 1H), 5.44-5.30 (m, 4H), 5.15 (d, J=7.0 Hz,1H), 5.05 (d, J=6.8 Hz, 1H), 4.07 (d, J=17.0 Hz, 1H), 3.90 (d, J=17.0Hz, 1H), 3.80 (d, J=7.0 Hz, 2H), 3.60-3.51 (m, 2H), 2.68-2.62 (m, 1H),2.50-2.47 (m, 1H), 2.38-2.29 (m, 2H), 2.14-2.02 (m, 2H), 1.25 (t, J=7.0Hz, 3H), 1.17 (t, J=7.0 Hz, 3H); ¹³C NMR (100 MHz, CDCl₃, 25° C.):δ=195.7, 166.7, 154.8, 154.2, 147.3, 140.7, 133.3, 132.1, 128.5, 128.3(×2), 128.2, 127.9, 127.7, 126.7 (×2), 120.1, 118.1, 102.9, 93.9, 93.4,77.4, 64.8, 64.4, 44.5, 40.5, 30.7, 15.0, 14.9; HRMS (ESI-TOF): m/z:calculated for C₂₉H₃₃O₇ClNa: 551.1680, found 551.1807 [M+Na⁺]. [α]²⁵_(D)=−40.4 (c 0.79, CHCl₃).

Compound (S)-2-112a: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=7.53-7.51 (m,2H), 7.44-7.33 (m, 3H), 7.14 (s, 1H), 6.88-6.81 (m, 1H), 6.02 (d, J=15.2Hz, 1H), 5.78 (dd, J=10.5, 1.8 Hz, 1H), 5.46-5.33 (m, 4H), 5.19 (d,J=7.0 Hz, 1H), 5.09 (d, J=7.6 Hz, 1H), 4.10 (d, J=17.0 Hz, 1H), 3.94 (d,J=17.5 Hz, 1H), 3.83 (d, J=7.0 Hz, 2H), 3.64-3.55 (m, 2H), 2.70-2.64 (m,1H), 2.54-2.50 (m, 1H), 2.37-2.33 (m, 2H), 2.15-2.08 (m, 2H), 1.29 (t,J=7.3 Hz, 3H), 1.21 (t, J=7.0 Hz, 3H); ¹³C NMR (100 MHz, CDCl₃, 25° C.):δ=195.7, 166.7, 154.8, 154.2, 147.3, 140.7, 133.2, 132.1, 128.5, 128.2,127.9, 127.7, 126.7, 120.1, 118.0, 102.8, 93.9, 93.5, 77.4, 64.8, 64.4,44.5, 40.5, 30.7, 15.0, 14.9, (one carbon is not detected); HRMS(ESI-TOF): m/z: calculated for C₂₉H₃₃O₇ClNa: 551.1680, found 551.1704[M+Na⁺]. [α]²⁵ _(D)=+48.8 (c 1.00, CHCl₃).

Compound (R)-2-112d: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=7.14 (s, 1H),6.72-6.66 (m, 1H), 5.88 (d, J=15.2 Hz, 1H), 5.33-5.17 (m, 6H), 4.92-4.88(m, 1H), 4.21 (d, J=17.0 Hz, 1H), 3.92 (d, J=17.0 Hz, 1H), 3.79-3.67 (m,4H), 2.33-2.17 (m, 5H), 2.07-1.96 (m, 2H), 1.23 (t, J=7.0 Hz, 3H), 1.21(t, J=7.0 Hz, 3H), 1.00 (d, J=5.8 Hz, 6H); ¹³C NMR (100 MHz, CDCl₃, 25°C.): δ=195.7, 167.1, 154.7, 154.4, 147.4, 133.7, 131.2, 128.8, 128.4,119.7, 118.0, 102.7, 93.9, 93.5, 80.0, 64.8, 64.5, 44.1, 32.3, 31.2,30.7, 30.6, 18.3, 17.2, 15.0, 14.9; HRMS (ESI-TOF): m/z: calculated forC₂₆H₃₅O₇ClNa: 517.1964, found 517.1844 [M+Na⁺]. [α]²⁵ _(D)=+21.3 (c1.00, CHCl₃).

Compound 2-112g: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=7.12 (s, 1H),6.76-6.70 (m, 1H), 5.87 (d, J=15.0 Hz, 1H), 5.33 (s, 2H), 5.26 (s, 2H),5.24-5.16 (m, 2H), 4.25 (t, J=5.1 Hz, 2H), 3.82-3.73 (m, 6H), 2.40-2.36(m, 2H), 2.16-2.13 (m, 4H), 1.27-1.23 (m, 6H); ¹³C NMR (100 MHz, CDCl₃,25° C.): δ=195.2, 167.2, 154.6, 153.4, 146.8, 132.3, 131.3, 128.9,128.5, 121.0, 117.8, 103.0, 94.0, 93.7, 64.8, 64.7, 64.6, 45.4, 31.9,31.0, 30.7, 15.0, 15.0; HRMS (ESI-TOF): m/z: calculated for C₂₃H₃₀O₇Cl:453.1675, found 453.1672 [M+H⁺].

Compound (S)-2-120: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=6.81-6.72 (m,1H), 6.74 (d, J=1.8 Hz, 1H), 6.56 (d, J=1.8 Hz, 1H), 5.98 (d, J=15.8 Hz,1H), 5.38-5.33 (m, 2H), 5.22-5.13 (m, 5H), 4.06 (d, J=14.6 Hz, 1H),3.75-3.65 (m, 4H), 3.46 (d, J=14.6 Hz, 1H), 2.37-2.22 (m, 4H), 2.18-2.02(m, 2H), 1.39 (d, J=5.8 Hz, 3H), 1.22 (t, J=7.0 Hz, 3H), 1.18 (t, J=7.0Hz, 3H); ¹³C NMR (100 MHz, CDCl₃, 25° C.): δ=197.4, 167.7, 159.0, 156.1,148.9, 135.0, 131.7, 129.8, 128.5, 118.5, 109.7, 102.2, 93.4, 93.0,71.5, 64.5, 64.4, 44.3, 39.5, 30.9, 30.6, 20.2, 15.0 (×2); HRMS(ESI-TOF): m/z: calculated for C₂₄H₃₂O₇Na: 455.2040, found 455.1901[M+Na⁺]. (+)-(2S): [α]²⁵ _(D)=+59.5 (c 1.00, CHCl₃).

Compound (R)-2-120a: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=7.56-7.54 (m,2H), 7.41-7.29 (m, 3H), 6.89-6.82 (m, 1H), 6.78 (d, J=2.3 Hz, 1H), 6.61(d, J=1.8 Hz, 1H), 6.06 (d, J=16.4 Hz, 1H), 5.98 (dd, J=11.7, 2.4 Hz,1H), 5.53-5.51 (m, 2H), 5.20 (d, J=7.0 Hz, 1H), 5.17 (d, J=6.4 Hz, 1H),5.07 (d, J=7.0 Hz, 1H), 4.96 (d, J=7.0 Hz, 1H), 4.20 (d, J=14.6 Hz, 1H),3.73-3.68 (m, 2H), 3.54-3.45 (m, 3H), 2.71-2.66 (m, 1H), 2.55-2.51 (m,1H), 2.38-2.32 (m, 2H), 2.23-2.06 (m, 2H), 1.22 (t, J=7.0 Hz, 3H), 1.14(t, J=7.0 Hz, 3H); ¹³C NMR (100 MHz, CDCl₃, 25° C.): δ=197.6, 167.4,159.3, 156.6, 149.0, 140.8, 135.6, 132.2, 129.9, 128.5, 128.2 (×2),127.9, 126.9 (×2), 117.9, 109.9, 102.3, 93.2, 93.0, 76.6, 64.4, 64.3,44.4, 40.5, 31.0, 30.6, 15.0, 14.9; HRMS (ESI-TOF): m/z: calculated forC₂₉H₃₄O₇Na: 517.2197, found 517.2062 [M+Na⁺]. [α]²⁵ _(D)=−108.3 (c 1.00,CHCl₃).

Compound (S)-2-120a: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=7.56-7.54 (m,2H), 7.41-7.29 (m, 3H), 6.90-6.82 (m, 1H), 6.78 (d, J=2.4 Hz, 1H), 6.61(d, J=1.8 Hz, 1H), 6.07 (d, J=16.4 Hz, 1H), 5.98 (dd, J=11.4, 2.0 Hz,1H), 5.53-5.51 (m, 2H), 5.20 (d, J=7.0 Hz, 1H), 5.18 (d, J=7.0 Hz, 1H),5.07 (d, J=7.0 Hz, 1H), 4.97 (d, J=7.0 Hz, 1H), 4.20 (d, J=14.6 Hz, 1H),3.74-3.69 (m, 2H), 3.55-3.46 (m, 3H), 2.71-2.66 (m, 1H), 2.55-2.52 (m,1H), 2.38-2.33 (m, 2H), 2.23-2.09 (m, 2H), 1.22 (t, J=7.0 Hz, 3H), 1.15(t, J=7.0 Hz, 3H); ¹³C NMR (100 MHz, CDCl₃, 25° C.): δ=197.6, 167.5,159.3, 156.6, 149.0, 140.8, 135.6, 132.2, 129.9, 128.5, 128.2 (×2),127.9, 126.9 (×2), 117.9, 110.0, 102.3, 93.2, 93.0, 76.6, 64.4, 64.3,44.4, 40.5, 31.0, 30.6, 15.0, 14.9; HRMS (ESI-TOF): m/z: calculated forC₂₉H₃₄O₇Na: 517.2197, found 517.2049 [M+Na⁺]. [α]²⁵ _(D)=+81.6 (c 1.00,CHCl₃).

Compound (R)-2-120d: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=6.83 (d, J=1.7Hz, 1H), 6.79-6.72 (m, 1H), 6.61 (d, J=1.2 Hz, 1H), 6.00 (d, J=16.4 Hz,1H), 5.38-5.36 (m, 2H), 5.26-5.09 (m, 5H), 4.30 (d, J=14.6 Hz, 1H),3.74-3.67 (m, 4H), 3.46 (d, J=14.6 Hz, 1H), 2.33-2.26 (m, 4H), 2.18-2.14(m, 2H), 2.06-2.01 (m, 1H), 1.23 (t, J=7.0 Hz, 3H), 1.21 (t, J=7.0 Hz,3H), 1.06 (d, J=6.6 Hz, 3H), 1.05 (d, J=7.0 Hz, 3H); ¹³C NMR (100 MHz,CDCl₃, 25° C.): δ=197.9, 167.6, 159.2, 156.9, 149.2, 136.4, 131.5,129.9, 129.2, 117.4, 109.8, 102.0, 93.3, 93.0, 78.9, 64.4 (×2), 44.2,33.0, 32.0, 31.0, 30.4, 18.3, 17.2, 15.0 (×2); HRMS (ESI-TOF): m/z:calculated for C₂₆H₃₆O₇Na: 483.2353, found 483.2215 [M+Na⁺]. [α]²⁵_(D)=+52.8 (c 1.00, CHCl₃).

Compound (R)-2-120e: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=6.77 (d, J=1.8Hz, 1H), 6.77-6.70 (m, 1H), 6.57 (d, J=1.7 Hz, 1H), 5.97 (d, J=16.4 Hz,1H), 5.37-5.32 (m, 2H), 5.21-5.14 (m, 5H), 4.17 (d, J=14.6 Hz, 1H),3.73-3.65 (m, 4H), 3.45 (d, J=14.6 Hz, 1H), 2.39-2.20 (m, 4H), 2.17-2.00(m, 2H), 1.78-1.72 (m, 1H), 1.69-1.60 (m, 1H), 1.54-1.44 (m, 2H), 1.22(t, J=7.0 Hz, 3H), 1.18 (t, J=7.0 Hz, 3H), 0.97 (d, J=7.3 Hz, 3H); ¹³CNMR (100 MHz, CDCl₃, 25° C.): δ=197.7, 167.6, 159.1, 156.6, 149.0,135.8, 131.5, 129.9, 128.7, 117.9, 109.8, 102.1, 93.3, 93.0, 74.5, 64.4,64.3, 44.2, 37.3, 37.0, 31.0, 30.5, 18.2, 15.0, 14.9, 14.2. [α]²⁵_(D)=−1.3 (c 1.00, CHCl₃).

Compound (R)-2-120f: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=7.39-7.33 (m,4H), 7.31-7.27 (m, 1H), 6.82 (s, 1H), 6.82-6.75 (m, 1H), 6.63 (s, 1H),6.02 (d, J=16.4 Hz, 1H), 5.35-5.29 (m, 2H), 5.27-5.20 (m, 5H), 4.16 (d,J=14.6 Hz, 1H), 3.79-3.70 (m, 4H), 3.52 (d, J=14.6 Hz, 1H), 3.37 (dd,J=13.4, 4.1 Hz, 1H), 2.78 (dd, J=13.5, 9.4 Hz, 1H), 2.37-2.12 (m, 5H),2.06-2.02 (m, 1H), 1.26 (t, J=7.0 Hz, 3H), 1.24 (t, J=7.0 Hz, 3H); ¹³CNMR (100 MHz, CDCl₃, 25° C.): δ=197.6, 167.8, 159.2, 156.5, 149.0,137.3, 135.5, 131.8, 129.9, 129.5 (×2), 128.6 (×2), 128.4, 126.7, 118.1,109.9, 102.3, 93.5, 93.1, 75.8, 64.6, 64.4, 44.4, 41.0, 36.2, 31.0,30.6, 15.0 (×2); HRMS (ESI-TOF): m/z: calculated for C₃₀H₃₆O₇Na:531.2359, found 531.2350 [M+Na⁺]. [α]²⁵ _(D)=−24.1 (c 0.33, CHCl₃).

Compound 2-120g: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=6.85-6.78 (m+s, 2H),6.56 (d, J=2.1 Hz, 1H), 6.00 (d, J=16.1 Hz, 1H), 5.38-5.34 (m, 2H), 5.23(s, 2H), 5.19 (s, 2H), 4.33 (t, J=5.4 Hz, 2H), 3.75-3.70 (m, 6H),2.45-2.41 (m, 2H), 2.19 (bs, 4H), 1.27-1.20 (m, 6H); ¹³C NMR (100 MHz,CDCl₃, 25° C.): δ=197.2, 168.1, 159.1, 155.8, 148.6, 134.5, 131.7,129.8, 129.1, 118.9, 109.9, 102.4, 93.6, 93.1, 64.5, 64.5, 64.4, 45.4,31.9, 31.1, 30.8, 15.0, 15.0; HRMS (ESI-TOF): m/z: calculated forC₂₃H₃₀O₇Na: 441.1884, found 441.1888 [M+Na⁺].

Compound (S)-2-121: Mixture of four diastereoisomers; ¹H NMR (400 MHz,CDCl₃, 25° C.): δ=6.77-6.72 (m, 1H), 6.57-6.47 (m, 1H), 5.62-5.35 (m,3H), 5.24-5.16 (m, 4H), 3.77-3.70 (m, 4H), 3.62-3.60 (m, 1.5H),3.53-3.49 (m, 1.5H), 3.17-3.11 (m, 3H), 3.04-2.97 (m, 1H), 2.57-2.44 (m,2H), 2.36-1.99 (m, 6H), 1.37-1.33 (m, 3H), 1.28-1.21 (m, 8H); HRMS(ESI-TOF): m/z: calculated for C₂₆H₃₉O₈NNa: 516.2568, found 516.2596[M+Na⁺].

Compound (R)-2-121a: Mixture of four diastereoisomers; ¹H NMR (400 MHz,CDCl₃, 25° C.): δ=7.51-7.42 (m, 2H), 7.38-7.31 (m, 3H), 6.73-6.70 (m,1H), 6.60-6.49 (m, 1H), 6.45-6.31 (m, 1H), 5.73-5.39 (m, 2H), 5.23-5.00(m, 4H), 3.75-3.69 (m, 2H), 3.56-3.34 (m, 6H), 3.19-3.09 (m, 3H),2.66-2.08 (m, 8H), 1.31-1.19 (m, 5H), 1.10-1.04 (m, 3H); HRMS (ESI-TOF):m/z: calculated for C₃₁H₄₁O₈NNa: 578.2724, found 578.2715 [M+Na⁺].

Compound (S)-2-121a: Mixture of four diastereoisomers; ¹H NMR (400 MHz,CDCl₃, 25° C.): δ=7.51-7.42 (m, 2H), 7.38-7.29 (m, 3H), 6.73-6.70 (m,1H), 6.60-6.49 (m, 1H), 6.45-6.31 (m, 1H), 5.73-5.42 (m, 2H), 5.25-5.01(m, 4H), 3.76-3.69 (m, 2H), 3.62-3.34 (m, 6H), 3.20-3.09 (m, 3H),2.66-2.50 (m, 2H), 2.22-2.12 (m, 4H), 1.72-1.66 (m, 2H), 1.31-1.19 (m,5H), 1.10-1.04 (m, 3H); HRMS (ESI-TOF): m/z: calculated for C₃₁H₄₁O₈NNa:578.2724, found 578.2720 [M+Na⁺].

Compound (R)-2-121d: Mixture of four diastereoisomers; ¹H NMR (400 MHz,CDCl₃, 25° C.): δ=6.77 (s, 1H), 6.52 (s, 0.5H), 6.46 (s, 0.5H),5.59-5.37 (m, 2H), 5.21-5.18 (m, 4H), 5.09-4.92 (m, 1H), 3.75-3.70 (m,4H), 3.53-3.48 (m, 3H), 3.38-3.34 (m, 1H), 3.19-3.10 (m, 3H), 2.65-2.47(m, 3H), 2.29-2.04 (m, 6H), 1.89-1.72 (m, 2H), 1.31-1.20 (m, 6H),1.06-0.96 (m, 6H); HRMS (ESI-TOF): m/z: calculated for C₂₈H₄₃O₈NNa:544.2881, found 544.2907 [M+Na⁺].

Compound 2-121g: Mixture of two diastereoisomers; ¹H NMR (400 MHz,CDCl₃, 25° C.): 6=6.74-6.72 (m, 1H), 6.54 (d, J=1.8 Hz, 0.6H), 6.50 (d,J=1.7 Hz, 0.4H), 5.52-5.41 (m, 2H), 5.21 (s, 2H), 5.19 (s, 2H),4.69-4.65 (m, 1H), 4.58-4.47 (m, 1H), 3.74-3.68 (m, 4H), 3.55 (s, 3H),3.13 (s, 3H), 2.97-2.94 (m, 1H), 2.52-2.40 (m, 2H), 2.26-1.98 (m, 6H),1.72-1.58 (m, 2H), 1.24-1.20 (m, 6H); HRMS (ESI-TOF): m/z: calculatedfor C₂₅H₃₈O₈N: 480.2567, found 480.2592 [M+H⁺].

General procedure for the EOM deprotection to generate compoundsdeprotected-2-121a-g, 2-85a-g and 2-103a-g: PS-TsOH (10.0 equiv., 3.2mmol.g⁻¹) was added to a solution of the corresponding compound 2-121a-gor 2-112a-g or 2-120a-g (1.0 equiv.) in MeOH (0.03 M) and the resultingsuspension was shaken at 40° C. for 1 to 4 h. After this time, thereaction mixture was filtered and the methanolic solution concentratedunder reduced pressure. Purification by flash chromatography (silicagel, 0-33% EtOAc/cyclohexane gradient) afforded the correspondingcompound deprotected-2-121a-g or 2-85a-g or 2-103a-g (>90%).

Deprotected compound (S)-2-121: Mixture of four diastereoisomers; ¹H NMR(400 MHz, CDCl₃, 25° C.): δ=12.90 (s, 0.5H), 12.83 (s, 0.5H), 12.12 (s,0.5H), 12.02 (s, 0.5H), 6.92 (s, 0.5H), 6.84 (s, 0.5H), 6.83 (s, 1H),6.79 (s, 0.5H), 6.60 (s, 1H), 6.54 (s, 0.5H), 5.60-5.29 (m, 4H),5.17-4.99 (m, 2H), 4.14-3.99 (m, 2H), 2.98-2.72 (m, 12H), 2.60-1.92 (m,16H), 1.31 (d, J=6.4 Hz, 1.5H), 1.22 (d, J=8.7 Hz, 1.5H), 1.13-1.02 (m,7H); HRMS (ESI-TOF): m/z: calculated for C₂₀H₂₇O₆NNa: 400.1851, found400.1731 [M+Na⁺].

Deprotected compound (R)-2-121a: Mixture of four diastereoisomers; ¹HNMR (400 MHz, CDCl₃, 25° C.): δ=11.90 (s, 0.25H), 11.08 (s, 0.5H), 10.98(s, 0.25H), 7.38-7.29 (m, 5H), 6.39 (s, 0.25H), 6.33 (s, 0.25H), 6.29(s, 1.25H), 6.26 (s, 0.25H), 6.05-5.95 (m, 1H), 5.70-5.52 (m, 2H),4.18-4.03 (m, 1H), 3.51-3.49 (m, 3H), 3.16-3.14 (m, 3H), 2.75-2.62 (m,2H), 2.36-2.29 (m, 2H), 2.12-1.96 (m, 4H), 1.81-1.73 (m, 2H),para-phenol not detected; HRMS (ESI-TOF): m/z: calculated for C₂₅H₃₀O₆N,440.2068. found 440.2103 [M+H⁺].

Deprotected compound (S)-2-121a: Mixture of four diastereoisomers; ¹HNMR (400 MHz, CDCl₃, 25° C.): δ=11.90 (s, 0.25H), 11.08 (s, 0.5H), 10.98(s, 0.25H), 7.38-7.31 (m, 5H), 6.38 (s, 0.25H), 6.33 (s, 0.25H), 6.29(s, 1.25H), 6.26 (s, 0.25H), 6.05-5.95 (m, 1H), 5.71-5.54 (m, 2H),4.13-4.04 (m, 1H), 3.53-3.50 (m, 3H), 3.19-3.14 (m, 3H), 2.78-2.63 (m,2H), 2.33-2.29 (m, 2H), 2.16-2.04 (m, 4H), 1.81-1.68 (m, 2H),para-phenol not detected; HRMS (ESI-TOF): m/z: calculated forC₂₅H₂₉O₆NNa: 462.1887, found 462.2080 [M+Na⁺].

Deprotected compound (R)-2-121d: Mixture of four diastereoisomers; ¹HNMR (400 MHz, CDCl₃, 25° C.): δ=11.54 (s, 1H), 6.33 (d, J=2.3 Hz, 1H),6.25 (s, 1H), 5.53-5.51 (m, 1H), 5.44-5.41 (m, 1H), 5.11-5.08 (m, 1H),4.01 (d, J=11.7 Hz, 2H), 3.45 (s, 3H), 3.11 (s, 3H), 2.83-2.73 (m, 1H),2.68-2.59 (m, 1H), 2.27-2.20 (m, 1H), 2.10-1.87 (m, 6H), 1.82-1.72 (m,1H), 1.01-0.94 (m, 6H), para-phenol not detected; HRMS (ESI-TOF): m/z:calculated for C₂₂H₃₁O₆NNa: 428.2044, found 428.2109 [M+Na⁺].

Deprotected compound 2-121g: Mixture of two diastereoisomers; ¹H NMR(400 MHz, CDCl₃, 25° C.): δ=12.33 (s, 0.5H), 11.85 (s, 0.5H), 6.34-6.32(m, 1H), 6.25-6.22 (m, 1H), 5.62-5.45 (m, 2H), 4.54-4.37 (m, 1H),4.29-4.21 (m, 1H), 3.53-3.49 (m, 3H), 3.15-3.12 (m, 3.5H), 2.95-2.86 (m,0.5H), 2.67-2.52 (m, 2H), 2.39-1.96 (m, 8H), para-phenol not detected;HRMS (ESI-TOF): m/z: calculated for C₁₉H₂₆O₆N, 364.1755. found 364.1715[M+H⁺].

Compound (S)-2-85: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=12.40 (s, 1H), 6.83(s, 1H), 6.66-6.61 (m, 1H), 5.96 (bs, 1H), 5.80 (d, J=15.2 Hz, 1H),5.16-5.12 (m, 1H), 4.99-4.91 (m, 1H), 4.75-4.68 (m, 1H), 4.26 (d, J=17.5Hz, 1H), 4.13 (d, J=17.5 Hz, 1H), 2.52-2.45 (m, 1H), 1.86-1.79 (m, 3H),1.75-1.67 (m, 1H), 1.54-1.49 (m, 1H), 0.97 (d, J=7.0 Hz, 3H); ¹³C NMR(100 MHz, C₆D₆, 25° C.): δ=193.8, 169.8, 164.3, 156.8, 146.0, 137.1,131.9, 128.1, 126.2, 115.2, 107.6, 103.6, 72.4, 46.2, 36.3, 31.0, 30.8,17.2. (−)-(2S): [α]²⁵ _(D)=−21.9 (c 0.62, CHCl₃).

Compound (R)-2-85a: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=12.25 (bs, 1H),7.32-7.29 (m, 2H), 7.19-7.15 (m, 3H), 6.80 (s, 1H), 6.81-6.75 (m, 1H),6.29-6.26 (m, 1H), 5.92 (d, J=15.8 Hz, 1H), 5.80 (s, 1H), 5.05-4.99 (m,1H), 4.81-4.75 (m, 1H), 4.56 (d, J=17.6 Hz, 1H), 4.12 (d, J=17.5 Hz,1H), 2.73-2.66 (m, 1H), 2.39-2.35 (m, 1H), 1.86-1.76 (m, 2H), 1.64-1.49(m, 2H); ¹³C NMR (100 MHz, C₆D₆, 25° C.): δ=194.2, 169.8, 164.4, 156.8,145.5, 138.3, 137.0, 132.9, 129.9 (×2), 128.6, 127.3, 126.6 (×2), 125.8,115.2, 107.5, 103.6, 77.6, 46.6, 38.3, 31.0, 30.6; HRMS (ESI-TOF): m/z:calculated for C₂₃H₂₁O₅ClNa: 435.0970, found 435.0914 [M+Na⁺]. [α]²⁵_(D)=−12.0 (c 0.55, CHCl₃).

Compound (S)-2-85a: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=12.20 (bs, 1H),7.15-7.09 (m, 3H), 6.75 (s, 1H), 6.77-6.71 (m, 1H), 6.25-6.22 (m, 1H),5.88 (d, J=15.2 Hz, 1H), 5.72 (s, 1H), 5.00-4.95 (m, 1H), 4.77-4.73 (m,1H), 4.52 (d, J=17.6 Hz, 1H), 4.08 (d, J=17.5 Hz, 1H), 2.68-2.62 (m,1H), 2.35-2.31 (m, 1H), 1.81-1.78 (m, 2H), 1.56-1.49 (m, 2H), 2H maskedby the solvent peak; ¹³C NMR (100 MHz, C₆D₆, 25° C.): δ=194.2, 169.8,164.4, 156.8, 145.5, 138.3, 137.0, 132.2, 129.9 (×2), 128.5, 127.3,126.5 (×2), 125.8, 115.2, 107.5, 103.6, 77.5, 46.6, 38.2, 31.0, 30.6;HRMS (ESI-TOF): m/z: calculated for C₂₃H₂₁O₅ClNa: 435.0970, found435.0885 [M+Na⁺]. [α]²⁵ _(D)=+11.6 (c 0.51, CHCl₃).

Compound (R)-2-85d: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=12.31 (s, 1H),6.83 (s, 1H), 6.74-6.67 (m, 1H), 5.84 (bs, 1H), 5.82 (d, J=15.8 Hz, 1H),5.03-4.95 (m, 1H), 4.88-4.86 (m, 1H), 4.76-4.70 (m, 1H), 4.40 (d, J=17.6Hz, 1H), 4.15 (d, J=17.5 Hz, 1H), 2.40-2.34 (m, 1H), 2.22-2.18 (m, 1H),1.87-1.65 (m, 4H), 1.53-1.48 (m, 1H), 0.92 (d, J=6.4 Hz, 3H), 0.66 (d,J=7.0 Hz, 3H); ¹³C NMR (100 MHz, C₆D₆, 25° C.): δ=193.7, 164.2, 156.8,145.8, 137.2, 131.8, 129.3, 126.3, 115.3, 107.9, 103.6, 82.1, 46.4,33.3, 30.9, 30.7, 28.8, 20.1, 18.5, 18.3; HRMS (ESI-TOF): nm/z:calculated for C₂₀H₂₃ClO₅Na: 401.1126, found 401.1170 [M+Na⁺]. [α]²⁵_(D)=−35.6 (c 0.52, CHCl₃).

Compound 2-85g: ¹H NMR (400 MHz, CD₃OD, 25° C.): δ=6.74-6.68 (m, 1H),6.48 (s, 1H), 5.86 (d, J=15.2 Hz, 1H), 5.31-5.25 (m, 2H), 4.39 (t, J=5.3Hz, 2H), 4.27 (s, 2H), 2.43-2.40 (m, 2H), 2.25 (m, 4H), phenols notdetected; ¹³C NMR (100 MHz, CD₃OD, 25° C.): δ=196.9, 170.1, 161.9,158.1, 147.8, 135.9, 130.9, 130.2, 129.9, 115.2, 107.3, 102.4, 65.9,46.2, 31.3, 30.9, 30.5; HRMS (ESI-TOF): m/z: calculated for C₁₇H₁₈O₅Cl:337.0837, found 337.0797 [M+H⁺].

Compound (S)-2-103: ¹H NMR (400 MHz, CD₃OD, 25° C.): δ=6.78-6.71 (m,1H), 6.29 (d, J=2.4 Hz, 1H), 6.22 (d, J=2.0 Hz, 1H), 5.87 (d, J=15.5 Hz,1H), 5.37-5.23 (m, 3H), 4.01 (d, J=17.2 Hz, 1H), 3.92 (d, J=17.0 Hz,1H), 2.67-2.61 (m, 1H), 2.29-2.15 (m, 5H), 1.31 (d, J=6.4 Hz, 3H),phenols not detected; ¹³C NMR (100 MHz, CD₃OD, 25° C.): δ=198.5, 169.8,164.2, 162.3, 148.4, 139.1, 131.6, 129.6, 127.3, 111.7, 101.7, 72.0,47.7, 36.8, 30.8, 30.7, 17.4, (one quartenary carbon is not detected);HRMS (ESI-TOF): m/z: calculated for C₁₈H₂₀O₅Na: 339.1203, found 339.1141[M+Na⁺]. (−)-(2S): [α]²⁵ _(D)=−45.1 (c 0.27, CHCl₃).

Compound (R)-2-103a: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=12.0 (bs, 1H),7.32-7.29 (m, 3H), 7.19-7.15 (m, 2H), 6.86-6.79 (m, 1H), 6.51 (d, J=2.4Hz, 1H), 6.27-6.25 (m, 1H), 6.11 (d, J=2.4 Hz, 1H), 6.02 (d, J=15.8 Hz,1H), 5.49 (s, 1H), 5.17-5.10 (m, 1H), 4.97-4.90 (m, 1H), 4.40 (d, J=16.4Hz, 1H), 3.97 (d, J=17.2 Hz, 1H), 2.83-2.76 (m, 1H), 2.45-2.38 (m, 1H),1.89-1.78 (m, 2H), 1.67-1.58 (m, 2H); ¹³C NMR (100 MHz, C₆D₆, 25° C.):δ=196.5, 169.6, 166.1, 161.3, 146.0, 140.5, 138.8, 132.1, 130.0, 128.6(×2), 127.3, 126.6 (×2), 126.3, 112.2, 105.9, 103.0, 77.1, 48.6, 38.4,30.9, 30.3; HRMS (ESI-TOF): m/z: calculated for C₂₃H₂₂O₅Na: 401.1359,found 401.1271 [M+Na⁺]. [α]²⁵ _(D)=−10.3 (c 0.25, CHCl₃).

Compound (S)-2-103a: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=12.0 (bs, 1H),7.27-7.21 (m, 3H), 7.17-7.13 (m, 2H), 6.87-6.79 (m, 1H), 6.55 (d, J=2.3Hz, 1H), 6.29-6.26 (m, 1H), 6.16 (d, J=2.3 Hz, 1H), 6.03 (d, J=15.8 Hz,1H), 5.74 (s, 1H), 5.18-5.12 (m, 1H), 4.98-4.91 (m, 1H), 4.41 (d, J=15.8Hz, 1H), 3.99 (d, J=16.9 Hz, 1H), 2.84-2.77 (m, 1H), 2.46-2.43 (m, 1H),1.85-1.79 (m, 2H), 1.70-1.58 (m, 2H); ¹³C NMR (100 MHz, C₆D₆, 25° C.):δ=196.9, 169.6, 166.2, 161.5, 146.3, 140.5, 138.9, 132.1, 130.0, 128.6(×2), 127.3, 126.6 (×2), 126.3, 112.2, 105.8, 103.0, 77.1, 48.6, 38.4,30.9, 30.4; HRMS (ESI-TOF): m/z: calculated for C₂₃H₂₂O₅Na: 401.1359,found 401.1264 [M+Na⁺]. [α]²⁵ _(D)=+11.9 (c 0.51, CHCl₃).

Compound (R)-2-103d: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=12.10 (s, 1H),6.79 (dt, J=15.2, 7.6 Hz, 1H), 6.59 (d, J=2.4 Hz, 1H), 6.22 (d, J=2.3Hz, 1H), 5.96 (d, J=15.8 Hz, 1H), 5.88 (bs, 1H), 5.13-5.05 (m, 1H),4.92-4.85 (m, 2H), 4.27 (d, J=15.8 Hz, 1H), 4.03 (d, J=15.8 Hz, 1H),2.50-2.44 (m, 1H), 2.24-2.20 (m, 1H), 1.96-1.71 (m, 4H), 1.63-1.56 (m,1H), 0.91 (d, J=6.3 Hz, 3H), 0.71 (d, J=6.4 Hz, 3H); ¹³C NMR (100 MHz,C₆D₆, 25° C.): δ=196.5, 169.7, 166.0, 161.4, 146.8, 140.8, 131.7, 129.5,126.9, 112.3, 106.0, 103.0, 81.3, 48.5, 33.7, 30.9, 30.4, 29.6, 19.7,18.4; HRMS (ESI-TOF): m/z: calculated for C₂₀H₂₄O₅Na: 367.1516, found367.1424 [M+Na⁺]. [α]²⁵ _(D)=−31.9 (c 0.50, CHCl₃).

Compound (R)-2-103e: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=12.43 (s, 1H),6.74 (d, J=1.7 Hz, 1H), 6.73-6.65 (m, 1H), 6.48 (d, J=1.7 Hz, 1H), 5.92(d, J=15.8 Hz, 1H), 5.12-5.00 (m, 2H), 4.91-4.80 (m, 1H), 4.19 (d,J=17.0 Hz, 1H), 3.84 (d, J=16.4 Hz, 1H), 2.77 (m, 1H), 2.64-2.57 (m,1H), 2.01-1.97 (m, 1H), 1.89-1.70 (m, 3H), 1.61-1.56 (m, 2H), 1.30-1.21(m, 2H), 0.90 (t, J=6.7 Hz, 3H), para-phenol not detected; ¹³C NMR (100MHz, CDCl₃, 25° C.): δ=197.5, 169.9, 165.6, 160.6, 147.5, 140.2, 131.9,129.5, 127.0, 112.8, 106.1, 102.9, 76.2, 48.7, 35.7, 34.3, 31.1, 29.7,19.4, 13.8; HRMS (ESI-TOF): m/z: calculated for C₂₀H₂₅O₅: 345.1697,found 345.1739 [M+H⁺]. [α]²⁵ _(D)=+21.6 (c 0.36, CHCl₃).

Compound (R)-2-103f: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=12.31 (s, 1H),7.19-7.13 (m, 5H), 6.80-6.72 (m, 1H), 6.53 (d, J=1.8 Hz, 1H), 6.05 (s,1H), 5.89 (d, J=15.8 Hz, 1H), 5.47-5.44 (m, 1H), 5.11-5.05 (m, 1H),4.85-4.81 (m, 1H), 4.10 (d, J=17.0 Hz, 1H), 3.62 (d, J=17.0 Hz, 1H),2.89-2.84 (m, 1H), 2.67-2.60 (m, 2H), 2.08-2.04 (m, 1H), 1.90-1.69 (m,3H), 1.52-1.44 (m, 1H), para-phenol not detected; ¹³C NMR (100 MHz,CDCl₃, 25° C.): δ=197.7, 169.9, 165.7, 160.7, 147.4, 140.2, 137.2,132.2, 129.5, 128.8 (×2), 128.7 (×3), 126.8, 112.4, 105.9, 102.9, 48.9,38.5, 35.3, 31.1 (×2), 29.7; HRMS (ESI-TOF): m/z: calculated forC₂₄H₂₅O₅: 393.1697, found 393.1765 [M+H⁺]. [α]²⁵ _(D)=+25.4 (c 0.41,CHCl₃).

Compound 2-103g: ¹H NMR (400 MHz, CD₃OD, 25° C.): δ=6.75-6.69 (m, 1H),6.29 (s, 1H), 6.29 (d, J=2.3 Hz, 1H), 5.90 (d, J=15.8 Hz, 1H), 5.30-5.28(m, 2H), 4.39 (t, J=5.2 Hz, 2H), 4.02 (s, 2H), 2.45-2.41 (m, 2H),2.28-2.24 (m, 4H), phenols not detected; ¹³C NMR (100 MHz, CD₃OD, 25°C.): δ=198.4, 170.6, 164.9, 162.5, 148.1, 139.2, 130.7, 130.6, 130.2,112.2, 105.0, 101.6, 65.7, 47.7, 31.4, 30.9, 30.5; HRMS (ESI-TOF): m/z:calculated for C₁₇H₁₉O₅: 303.1227, found 303.1179 [M+H⁺].

General procedure for the synthesis of compounds 2-141: BER-resin(Borohydride on Amberlite, 1.0 equiv., 2.5 mmol.g⁻¹) was added to asolution of corresponding compound 2-112a-g or 2-120a-g (1.0 equiv.) inMeOH (0.03 M) at 0° C. and the reaction was stirred for 12 h. Thereaction mixture was then filtered and concentrated under reducedpressure. Purification by flash chromatography (silica gel, 0-20%EtOAc/cyclohexane gradient) afforded 2-141 (˜60% yield) as a mixture oftwo diastereoisomers 1:1.

Selected example of compounds 2-141: ¹H NMR (400 MHz, CDCl₃, 25° C.):δ=7.05 (s, 1H), 6.99 (s, 1H), 5.64-5.57 (m, 2H), 5.54-5.53 (m, 2H),5.49-5.35 (m, 7H), 5.31-5.28 (m, 4H), 5.24-5.16 (m, 4H), 5.13-5.08 (m,1H), 4.68 (m, 1H), 4.56 (m, 1H), 3.81-3.69 (m, 8H), 3.25 (dd, J=13.9,8.0 Hz, 1H), 3.19 (dd, J=13.7, 4.8 Hz, 1H), 3.11 (dd, J=13.5, 10.1 Hz,1H), 2.90 (dd, J=13.9, 5.1 Hz, 1H), 2.35 (m, 9H), 2.09-1.95 (m, 1H),1.80-1.70 (m, 2H), 1.39 (d, J=2.9 Hz, 3H), 1.37 (d, J=3.2 Hz, 3H), 1.24(2×q, J=6.9, 5.0 Hz, 12H); HRMS (ESI-TOF): m/z: calculated forC₂₄H₃₃ClO₇Na: 491.1807, found 491.1729 [M+Na⁺].

General procedure for the synthesis of compounds 2-142: PS-TsOH (10.0equiv., 3.2 mmol.g⁻¹) was added to a solution of the correspondingcompound 2-141 (1.0 equiv.) in MeOH (0.02 M) and the suspension wasshaken at 40° C. for 4 h. The reaction mixture was then filtered and themethanolic solution concentrated under reduced pressure. Purification bypreparative TLC (silica gel, 25% EtOAc/cyclohexane) afforded 2-142 (˜90%yield) as a mixture of two diastereoisomers 1:1.

Selected example of compounds 2-142: ¹H NMR (400 MHz, (CD₃)₂CO, 25° C.):δ=12.30 (s, 2H), 11.43 (s, 2H), 6.75 (s, 2H), 6.00 (bdd, J=6.4, 6.2 Hz,1H), 5.97 (bdd, J=6.4, 6.2 Hz, 1H), 5.97 (bd, J=6.7 Hz, 1H), 5.77 (bd,J=6.7 Hz, 1H), 5.57-5.48 (m, 4H), 5.18-5.14 (m, 2H), 3.38-3.28 (m, 3H),3.02 (dd, J=16.1, 10.5 Hz, 1H), 2.41-2.09 (m, 12H), 1.11 (d, J=6.2 Hz,6H), alcohols not detected; HRMS (ESI-TOF): m/z: calculated forC₁₈H₂₁ClO₅Na: 375.0970, found 375.1029 [M+Na⁺].

General procedure for the synthesis of compounds 2-143: Ac₂O (1.2equiv.), morpholinomethyl polystyrene (1.2 equiv., 3.2 mmol.g⁻¹) andDMAP (0.05 equiv.) were added to a solution of the correspondingcompound 2-141 (1.0 equiv.) in DMF (0.02 M) at 23° C. and the mixturewas stirred for 30 min, followed by TLC until consumption of thestarting material. The resin was then filtered and the organic phase wasconcentrated under reduced pressure. Purification by preparative TLC(silica gel, 20% EtOAc/cyclohexane) afforded corresponding compound2-143 (˜80% yield) as a mixture of two diastereoisomers 1:1.

Selected example of compounds 2-143: ¹H NMR (400 MHz, CDCl₃, 25° C.):δ=7.04 (s, 1H), 7.01 (s, 1H), 5.86 (dd, J=15.0, 6.9 Hz, 1H), 5.67 (dd,J=12.4, 6.2 Hz, 1H), 5.60-5.54 (m, 4H), 5.48 (dd, J=7.2, 7.2 Hz, 1H),5.41-5.34 (m, 3H), 5.32-5.30 (m, 4H), 5.28-5.23 (m, 2H), 5.21 (dd,J=11.0, 6.7 Hz, 2H), 5.17 (dd, J=11.8, 6.9 Hz, 2H), 3.81-3.69 (m, 8H),3.43 (dd, J=14.2, 7.5 Hz, 1H), 3.23-3.15 (m, 2H), 2.85 (dd, J=13.9, 5.4Hz, 1H), 2.30-2.17 (m, 8H), 2.12 (s, 3H), 2.06 (s, 3H), 2.00-1.95 (m,4H), 1.39 (2×d, J=5.6 Hz, 6H), 1.24 (m, 12H); HRMS (ESI-TOF): m/z:calculated for C₂₆H₃₅ClO₈Na: 533.1913, found 533.1864 [M+Na⁺].

General procedure for the synthesis of compounds 2-144: PS-TsOH (10.0equiv., 3.2 mmol.g⁻¹) was added to a solution of corresponding compound2-143 (1.0 equiv.) in MeOH (0.02 M) and the suspension was shaken at 40°C. for 4 h. After this time, the reaction mixture was filtered and themethanolic solution concentrated under reduced pressure. Purification bypreparative TLC (silica gel, 20% EtOAc/cyclohexane) afforded compound2-144 (˜60% yield).

Selected example of compounds 2-144: Mixture of diastereoisomers 2:1; ¹HNMR (400 MHz, CDCl₃, 25° C.): δ=12.60 (s, 1H), 12.12 (s, 0.5H), 6.93 (d,J=8.7 Hz, 0.5H), 6.66 (s, 1H), 6.64 (s, 0.5H), 6.62-6.60 (m, 1H),6.10-6.05 (m, 3H), 5.47-5.33 (m, 6H), 2.60-2.53 (m, 1.5H), 2.26-2.02 (m,7.5H), 1.44 (d, J=6.2 Hz, 1.5H), 1.43 (d, J=6.4 Hz, 3H), para-phenol notdetected; HRMS (ESI-TOF): m/z: calculated for C₁₈H₁₉ClO₄Na: 357.0864,found 357.0898 [M+Na⁺].

General procedure for the preparation of compounds 2-145: PS-TsOH (10.0equiv.) was added to a solution of corresponding compound 2-85a-g (1.0equiv.) in methanol (0.03 M) and the suspension was stirred for 15 h at40° C. The reaction was then filtered and the resin washed several timeswith CH₂Cl₂. Concentration under reduced pressure followed bypurification on preparative TLC (silica gel, 50% hexane/EtOAc) affordeddesired compound 2-145 as a mixture of diastereoisomers 2:1.

Selected example of compounds 2-145: ¹H NMR (400 MHz, C₆D₆, 25° C.):δ=12.28 (s, 0.4H), 11.91 (s, 0.6H), 7.21-7.11 (m, 5H), 6.62 (s, 1H),6.03-6.01 (m, 1H), 5.58 (bs, 1H), 5.38-5.33 (m, 1H), 5.27-5.20 (m, 1H),4.76 (d, J=17.5 Hz, 0.6H), 4.02 (d, J=17.0 Hz, 0.4H), 4.18 (d, J=18.1Hz, 0.6H), 4.09 (d, J=17.0 Hz, 0.4H), 3.87 (bs, 0.4H), 3.81 (bs, 0.6H),3.15 (s, 1.8H), 3.12 (s, 1.2H), 2.83-2.78 (m, 1H), 2.45-2.30 (m, 2H),2.18-2.16 (m, 1H), 2.02-1.97 (m, 2H), 1.79-1.72 (m, 2H); HRMS (ESI-TOF):m/z: calculated for C₂₄H₂₅O₆ClNa: 467.1232, found 467.1366 [M+Na⁺].

General procedure for the synthesis of compounds 2-146:(Polystyrylmethyl)trimethyl-ammonium cyanoborohydride (2.0 equiv., 3.5mmol.g⁻¹) was added to a solution of corresponding compound 2-85a-g or2-103a-g (1.0 equiv.) in CH₂Cl₂/AcOH 10:1 (0.08 M) at 23° C. and thereaction was monitored by TLC until consumption of the starting material(4 h). The resin was then filtered and the organic phase wasconcentrated under reduced pressure. Purification by preparative TLC(silica gel, 30% EtOAc/cyclohexane) afforded compound 2-146 (50-60%yield).

Selected example of compounds 2-146: ¹H NMR (400 MHz, CDCl₃, 25° C.):δ=11.75 (s, 1H), 6.65 (s, 1H), 5.48 (m, 2H), 5.49 (ddt, J=6.1, 3.5, 2.9Hz, 1H), 4.53 (d, J=17.5 Hz, 1H), 4.04 (d, J=17.7 Hz, 1H), 2.61-2.54 (m,2H), 2.48-2.28 (m, 3H), 2.19-2.14 (m, 1H), 2.08-1.99 (m, 1H), 1.72-1.61(m, 3H), 1.41 (d, J=6.4 Hz, 3H), para-phenol not detected; HRMS(ESI-TOF): m/z: calculated for C₁₈H₂₁ClO₅Na: 375.0970, found 375.1050[M+Na⁺].

General procedure for the synthesis of compounds 2-147: Thecorresponding alcohol R²OH (2.0 equiv.), triphenylphosphine (2.0 equiv.)and PS-DEAD (2.0 equiv., 1.3 mmol.g⁻¹) were added to a solution ofcorresponding compound 2-85a-g or 2-103a-g (1.0 equiv.) in THF (0.05 M)in a sequential manner. The reaction mixture was shaken at roomtemperature for 8 h, and then the resin was filtered and the filtrateswere directly purified by preparative TLC (silica gel, 10%EtOAc/cyclohexane) to afford a mixture of compound 2-147 along with thebis-alkylated product (78% yield).

Selected example of compounds 2-147: Mixture with the correspondingbis-allylated compound 1:1; ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=11.83 (s,1H), 6.82 (ddd, J=15.7, 8.2, 4.6 Hz, 1H), 6.72-6.65 (m, 1H), 6.46 (s,1H), 6.41 (s, 1H), 6.09-5.98 (m, 3H), 5.82 (d, J=15.7 Hz, 1H), 5.46-5.16(m, 8H), 4.57-4.54 (m, 3H), 4.51-4.49 (m, 3H), 4.19 (d, J=17.5 Hz, 1H),4.11 (d, J=14.6 Hz, 1H), 3.78 (d, J=17.0 Hz, 1H), 3.51 (d, J=14.2 Hz,1H), 2.76-2.69 (m, 1H), 2.38-2.05 (m, 11H), 1.42 (d, J=6.2 Hz, 3H), 1.35(d, J=6.3 Hz, 3H); mono-allylated compound: HRMS (ESI-TOF): m/z:calculated for C₂₁H₂₃ClO₅Na: 413.1132, found 413.1103 [M+Na⁺];bis-allylated compound: HRMS (ESI-TOF): m/z: calculated forC₂₄H₂₇ClO₅Na: 453.1449, found 453.1422 [M+Na⁺].

General procedure for the synthesis of compounds 2-148: TBD-methylpolystyrene (2.0 equiv., 2.9 mmol.g⁻¹) and the corresponding alkylbromide or chloride (BrCH₂COO^(t)Bu, EOMCl, 0.9 equiv.) were added to asolution of the corresponding compound 2-85a-g or 2-103a-g (1.0 equiv.)in CH₂Cl₂ (0.05 M) at 23° C. and the mixture was shaken for 3 h. Theresin was then filtered and the filtrates were concentrated underreduced pressure. Purification by preparative TLC (silica gel, 30%EtOAc/cyclohexane) afforded corresponding compound 2-148 (>90% yield).

Selected examples of compounds 2-148: ¹H NMR (400 MHz, CDCl₃, 25° C.):δ=11.84 (s, 1H), 6.69 (m, 1H), 6.41 (s, 1H), 5.76 (d, J=15.0 Hz, 1H),5.43 (m, 1H), 5.26 (ddd, J=15.0, 9.1, 4.8 Hz, 1H), 5.18-5.11 (m, 1H),4.65 (s, 2H), 4.33 (d, J=17.7 Hz, 1H), 4.16 (d, J=17.5 Hz, 1H),2.65-2.58 (m, 1H), 2.37-2.34 (m, 2H), 2.25-2.21 (m, 1H), 2.12-2.01 (m,2H), 1.53 (s, 9H), 1.34 (d, J=6.5 Hz, 3H); HRMS (ESI-TOF): m/z:calculated for C₂₄H₂₉ClO₇Na: 487.1494, found 487.1498 [M+Na⁺].

¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.76 (s, 1H), 6.86 (s, 1H), 6.70 (dt,J=14.9, 7.3 Hz, 1H), 5.77 (d, J=15.8 Hz, 1H), 5.46-5.42 (m, 1H), 5.37(s, 2H), 5.30-5.19 (m, 2H), 4.34 (d, J=17.6 Hz, 1H), 4.16 (d, J=18.1 Hz,1H), 3.80 (q, J=7.0 Hz, 2H), 2.66-2.59 (m, 1H), 2.37-2.34 (m, 2H),2.26-2.21 (m, 1H), 2.13-2.06 (m, 2H), 1.34 (d, J=6.4 Hz, 3H), 1.27 (t,J=7.0 Hz, 3H); HRMS (ESI-TOF): m/z: calculated for C₂₁H₂₅O₆ClNa:431.1237, found 431.1257 [M+Na⁺].

General procedure for the synthesis of compounds 2-149: OsO₄ (0.1equiv.) and NMO (1.0 equiv.) were added to a solution of compound2-85a-g or 2-103a-g (1.0 equiv.) in acetone/H₂O 10:1 (0.05 M) at 23° C.and the mixture was stirred for 1 h. The crude mixture was filteredthrough a pad of silica, concentrated and purified by preparative TLC(silica gel, 30% EtOAc/cyclohexane) to afford 2-149 as a mixture of twodiastereoisomers 1:1 (>70% yield).

Selected example of compounds 2-149: ¹H NMR (400 MHz, CD₃OD, 25° C.):δ=7.19 (m, 1H), 6.89-6.81 (m, 1H), 6.52 (s, 1H), 6.47 (s, 1H), 6.20 (d,J=16.1 Hz, 1H), 6.04 (d, J=15.6 Hz, 1H), 5.54-5.49 (m, 1H), 5.43-5.36(m, 1H), 4.50 (d, J=17.7 Hz, 1H), 4.46 (d, J=17.7 Hz, 1H), 4.39 (d,J=17.2 Hz, 1H), 4.07 (d, J=17.2 Hz, 1H), 3.80-3.64 (m, 2H), 3.51-3.46(m, 2H), 2.62-2.58 (m, 2H), 2.39-2.30 (m, 2H), 2.27-2.18 (m, 2H),2.08-1.98 (m, 2H), 2.00-1.85 (m, 4H), 1.44 (d, J=6.4 Hz, 6H), phenolsand alcohols not detected; HRMS (ESI-TOF): m/z: calculated forC₁₈H₂₁ClO₇Na: 407.0868, found 407.1031 [M+Na⁺].

General procedure for the synthesis of compounds 2-150: Freshly preparedDMDO (1.2 equiv., 0.04 M in acetone) was added to a solution of compound2-85a-g or 2-103a-g (1.0 equiv.) in CH₃CN (0.03 M) at 0° C. and themixture was stirred for 30 min. After evaporation of the solvents underreduced pressure, purification by preparative TLC (silica gel, 30%EtOAc/cyclohexane) afforded epoxides 2-150 (>90% yield) as a mixture oftwo diastereoisomers (1:1 to 3:1).

Selected examples of compounds 2-150: ¹H NMR (400 MHz, C₆D₆, 25° C.):δ=11.85 (s, 1H), 6.94-6.87 (m, 2H), 6.69 (s, 1H), 6.65 (s, 1H), 6.24(bd, J=15.2 Hz, 2H), 6.12 (d, J=15.8 Hz, 1H), 5.41-5.37 (m, 1H),5.33-5.30 (m, 1H), 4.54 (bd, J=18.1 Hz, 2H), 4.52-4.48 (m, 1H),4.40-4.34 (m, 1H), 4.27 (d, J=17.5 Hz, 1H), 2.78-2.72 (m, 2H), 2.58-2.55(m, 4H), 2.47-2.28 (m, 5H), 2.07 (m, 2H), 1.92-1.86 (m, 3H), 1.51 (d,J=6.4 Hz, 3H), 1.35 (d, J=6.4 Hz, 3H), para-phenol not detected; HRMS(ESI-TOF): mm/z: calculated for C₁₈H₁₉O₆ClNa: 389.0762, found 389.0844[M+Na⁺].

¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.80 (2×s, 2H), 7.43-7.18 (m, 10H),7.03-6.95 (m, 2H), 6.69 (s, 1H), 6.61 (s, 1H), 6.30 (d, J=16.4 Hz, 1H),6.21 (d, J=15.8 Hz, 1H), 6.15-6.10 (m, 1H), 6.03 (d, J=11.1 Hz, 1H),4.84 (2×d, J=18.1 Hz, 2H), 4.41 (2×d, J=17.6 Hz, 2H), 2.68-2.60 (m, 4H),2.41-2.27 (m, 8H), 1.83-1.76 (m, 4H), para-phenol not detected; HRMS(ESI-TOF): m/z: calculated for C₂₃H₂₁O₆ClNa: 451.0919, found 451.1028[M+Na⁺].

¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.56 (2×s, 2H), 6.92-6.82 (m, 2H),6.71 (s, 1H), 6.67 (s, 1H), 6.20 (m, 3H), 6.06 (d, J=15.8 Hz, 1H), 5.11(bs, 1H), 5.94 (m, 1H), 4.46 (2×d, J=18.1 Hz, 2H), 4.20 (2×d, J=18.1 Hz,2H), 2.72-2.70 (m, 2H), 2.53-2.48 (m, 4H), 2.38-2.35 (m, 3H), 2.25-2.13(m, 5H), 1.84-1.77 (m, 2H), 1.05-1.01 (m, 6H), 0.91-0.88 (m, 3H),0.86-0.84 (m, 3H), para-phenol not detected; HRMS (ESI-TOF): m/z:calculated for C₂₀H₂₃O₆ClNa: 417.1075, found 417.1128 [M+Na⁺].

¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.98 (s, 1H), 6.91-6.83 (m, 1H), 6.43(d, J=2.3 Hz, 1H), 6.24 (d, J=2.4 Hz, 1H), 6.11 (d, J=15.8 Hz, 1H), 5.35(bs, 1H), 5.29 (m, 1H), 4.52 (d, J=17.5 Hz, 1H), 3.63 (d, J=17.5 Hz,1H), 2.77 (m, 2H), 2.57-2.52 (m, 2H), 2.46-2.27 (m, 2H), 2.14-2.10 (m,1H), 1.93-1.88 (m, 1H), 1.48 (d, J=6.4 Hz, 3H); other isomer: ¹H NMR(400 MHz, C₆D₆, 25° C.): δ=11.67 (s, 1H), 6.89-6.83 (m, 1H), 6.40 (d,J=2.4 Hz, 1H), 6.24 (d, J=2.9 Hz, 1H), 6.21 (d, J=16.4 Hz, 1H), 5.37(bs, 1H), 5.22 (m, 1H), 4.20 (d, J=17.0 Hz, 1H), 4.06 (d, J=17.0 Hz,1H), 2.74 (m, 2H), 2.57-2.20 (m, 4H), 1.80-1.76 (m, 1H), 1.68-1.60 (m,1H), 1.37 (d, J=6.4 Hz, 3H), para-phenol not detected; HRMS (ESI-TOF):m/z: calculated for C₁₈H₂₀O₆Na: 355.1152, found 355.1249 [M+Na⁺].

¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.98 (s, 1H), 6.91-6.83 (m, 1H), 6.43(d, J=2.3 Hz, 1H), 6.24 (d, J=2.4 Hz, 1H), 6.11 (d, J=15.8 Hz, 1H), 5.35(bs, 1H), 5.29 (m, 1H), 4.52 (d, J=17.5 Hz, 1H), 3.63 (d, J=17.5 Hz,1H), 2.77 (bs, 1H), 2.57-2.52 (m, 2H), 2.46-2.27 (m, 2H), 2.14-2.10 (m,1H), 1.93-1.88 (m, 1H), 1.48 (d, J=6.4 Hz, 3H).

Major isomer: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.90 (s, 1H), 7.41-7.23(m, 5H), 6.95-6.89 (m, 1H), 6.42 (d, J=2.8 Hz, 1H), 6.27 (d, J=2.9 Hz,1H), 6.20 (d, J=15.8 Hz, 1H), 6.13 (d, J=4.1 Hz, 1H), 5.51 (m, 1H), 4.79(d, J=17.5 Hz, 1H), 3.79 (d, J=17.0 Hz, 1H), 2.68-2.55 (m, 3H),2.44-2.25 (m, 4H), para-phenol not detected; HRMS (ESI-TOF): m/z:calculated for C₂₃H₂₂O₆Na: 417.1309, found 417.1399 [M+Na⁺].

¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.77 (s, 1H), 6.92-6.82 (m, 1H), 6.44(d, J=2.3 Hz, 1H), 6.28 (d, J=2.4 Hz, 1H), 6.09 (d, J=15.8 Hz, 1H), 5.40(bs, 1H), 4.92 (m, 1H), 4.50 (d, J=17.5 Hz, 1H), 3.61 (d, J=17.5 Hz,1H), 2.72-2.70 (m, 1H), 2.56-2.45 (m, 2H), 2.38-2.15 (m, 4H), 1.91-1.85(m, 1H), 1.05-1.01 (m, 6H), para-phenol not detected; other isomer: ¹HNMR (400 MHz, C₆D₆, 25° C.): δ=11.55 (s, 1H), 6.86-6.79 (m, 1H), 6.42(s, 1H), 6.29 (s, 1H), 6.20 (d, J=15.8 Hz, 1H), 5.40 (m, 1H), 5.16 (m,1H), 4.14 (s, 1H), 4.12 (s, 1H), 2.72-2.70 (m, 1H), 2.53-2.37 (m, 4H),2.18-2.10 (m, 2H), 1.92-1.86 (m, 1H), 0.91-0.85 (m, 6H), para-phenol notdetected; HRMS (ESI-TOF): m/z: calculated for C₂₀H₂₄O₆Na: 383.1465,found 383.1574 [M+Na⁺].

¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.87 (s, 1H), 6.90-6.82 (m, 1H), 6.43(s, 1H), 6.26 (s, 1H), 6.10 (d, J=15.2 Hz, 1H), 5.31 (bs, 1H), 5.18 (bs,1H), 4.46 (d, J=17.5 Hz, 1H), 3.60 (d, J=17.6 Hz, 1H), 2.74 (bs, 1H),2.57-2.38 (m, 3H), 2.32-2.22 (m, 1H), 2.08-1.82 (m, 2H), 1.73-1.67 (m,1H), 1.42-1.37 (m, 2H), 1.33-1.28 (m, 2H), 1.01 (t, J=7.3 Hz, 3H); otherisomer: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.72 (s, 1H), 6.86-6.80 (m,1H), 6.41 (s, 1H), 6.27 (s, 1H), 6.21 (d, J=16.4 Hz, 1H), 5.37 (m, 1H),5.22 (m, 1H), 4.25 (d, J=16.4 Hz, 1H), 3.96 (d, J=16.4 Hz, 1H), 2.74(bs, 1H), 2.60-2.37 (m, 4H), 1.87-1.78 (m, 2H), 1.70-1.58 (m, 3H),1.38-1.22 (m, 2H), 0.95 (t, J=7.3 Hz, 3H); HRMS (ESI-TOF): m/z:calculated for C₂₀H₂₄O₆Na: 383.1465, found 383.1492 [M+Na⁺].

Major isomer: ¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.94 (s, 1H), 7.36-7.28(m, 5H), 6.95-6.88 (m, 1H), 6.42 (s, 1H), 6.22 (s, 1H), 6.11 (d, J=15.8Hz, 1H), 5.47 (m, 1H), 5.41 (bs, 1H), 4.43 (d, J=17.5 Hz, 1H), 3.56 (d,J=17.6 Hz, 1H), 3.19 (dd, J=13.7, 6.0 Hz, 1H), 3.03 (dd, J=13.7, 7.9 Hz,1H), 2.87 (bs, 1H), 2.70-2.28 (m, 4H), 2.03-1.93 (m, 2H), para-phenolnot detected; HRMS (ESI-TOF): nm/z: calculated for C₂₄H₂₄O₆Na: 431.1465,found 431.1578 [M+Na⁺].

General procedure for the preparation of macrocycles 2-151: HCl_(conc.)(20.0 equiv.) was added to a solution of compound 2-120 (1.0 equiv.) indioxane (0.05 M) at 23° C., and the mixture was stirred for 3 h. Afterthis time, the reaction was filtered through a pad of silica, thesolvents evaporated under reduced pressure, and purification bypreparative TLC (silica gel, 30% EtOAc/cyclohexane) afforded compound2-151 (>75% yield) as a mixture of two diastereoisomers 1:1.

Selected examples of compounds 2-151: ¹H NMR (400 MHz, CDCl₃, 25° C.):δ=12.11 (s, 1H), 11.78 (s, 1H), 6.51 (s, 1H), 6.43 (s, 1H), 6.41 (d,J=2.4 Hz, 1H), 6.37 (d, J=2.7 Hz, 1H), 6.21 (d, J=2.4 Hz, 1H), 6.11 (d,J=2.4 Hz, 1H), 5.59-5.51 (m, 3H), 5.40-5.32 (m, 3H), 4.54 (d, J=17.2 Hz,1H), 4.42 (d, J=17.2 Hz, 1H), 3.60 (d, J=17.2 Hz, 1H), 3.45 (d, J=17.0Hz, 1H), 3.28 (dd, J=18.5, 9.4 Hz, 1H), 3.11 (dd, J=13.7, 6.2 Hz, 1H),3.07 (dd, J=13.4, 4.6 Hz, 1H), 2.76 (dd, J=19.0, 6.2 Hz, 1H), 2.62 (ddd,J=15.5, 8.8, 4.0 Hz, 1H), 2.54 (ddd, J=15.3, 6.2, 3.2 Hz, 1H), 2.40-2.26(m, 4H), 2.25-2.13 (m, 4H), 2.03-1.91 (m, 2H), 1.42 (d, J=6.4 Hz, 3H),1.40 (d, J=6.4 Hz, 3H), para-phenol not detected; HRMS (ESI-TOF): m/z:calculated for C₁₈H₂₁ClO₅Na: 375.0970, found 375.0928 [M+Na⁺].

¹H NMR (400 MHz, C₆D₆, 25° C.): δ=11.76 (s, 0.5H), 11.36 (s, 0.5H),7.40-7.29 (m, 5H), 6.65 (s, 0.5H), 6.62 (s, 0.5H), 6.18 (t, J=5.8 Hz,1H), 6.14 (s, 0.5H), 6.12 (s, 0.5H), 5.67-5.62 (m, 1H), 5.55-5.49 (m,1H), 4.93 (d, J=18.1 Hz, 0.5H), 4.80 (d, J=17.1 Hz, 0.5H), 4.58-4.56 (m,1H), 4.38 (d, J=18.1 Hz, 0.5H), 4.18 (d, J=17.1 Hz, 0.5H), 3.33-3.27 (m,1H), 3.10 (dd, J=18.4, 3.8 Hz, 0.5H), 2.84-2.68 (m, 2.5H), 2.42-2.32 (m,2H), 2.23-2.17 (m, 1H), 2.13-2.04 (m, 1H), para-phenol not detected;HRMS (ESI-TOF): m/z: calculated for C₂₃H₂₂O₅Cl₂Na: 471.0737, found471.0754 [M+Na⁺].

Elimination of β-Cl from compound 2-151: PS-TBD (51 mg, 2.6 mmol.g⁻¹)was added to a solution of compound 2-151 (95 mg, 270/mol) in CH₂Cl₂ (5mL) at 23° C., and the mixture was stirred for 8 h. After this time, thereaction was filtered, the solvents were evaporated under reducedpressure, and the remaining residue was purified by flash chromatography(silica gel, 0-30% EtOAc/cyclohexane gradient) to afford 2-103 (84 mg,98%).

General procedure for the synthesis of compounds 2-152: Compound 2-120aor 2-112a (1.0 equiv.) was dissolved in a 1:5 mixture of TFA/CH₂Cl₂ andstirred for 2 h at room temperature. Evaporation of the solvents;followed by flash chromatography (silica gel, 0-50% Et₂O/hexane),afforded compound 2-152 (˜70% yield).

Selected example of compounds 2-152: ¹H NMR (400 MHz, C₆D₆, 25° C.):δ=11.74 (s, 1H), 7.22-7.14 (m, 5H), 6.52 (s, 1H), 6.46 (dt, J=15.2, 7.3Hz, 1H), 6.28 (s, 1H), 5.89 (t, J=7.0 Hz, 1H), 5.76 (s, 1H), 5.59 (d,J=15.2 Hz, 1H), 5.37 (ddd, J=15.2, 6.9, 6.9 Hz, 1H), 5.24 (ddd, J=15.2,7.3, 7.0 Hz, 1H), 2.56-2.49 (m, 1H), 2.44-2.39 (m, 1H), 2.01-1.92 (m,4H); ¹³C NMR (100 MHz, C₆D₆, 25° C.): δ=164.8, 162.8, 158.7, 153.1,137.6, 136.9, 136.1, 133.9, 128.7 (×2), 126.5 (×2), 124.2, 122.0, 102.4,100.8, 79.9, 39.0, 32.3, 31.4, four quaternary carbons are not detected;HRMS (ESI-TOF): m/z: calculated for C₂₅H₂₀ClF₃O₆Na: 531.0793, found531.0992 [M+Na⁺].

Macrocycle 2-153: DHP (3.7 μL, 40.8 μmol) and PS-TsOH (12.7 mg, 40.8μmol, 3.2 mmol.g⁻¹) were added to a solution of compound 2-103 (12.9 mg,40.8 μmol) in CH₂Cl₂ (1 mL) at 23° C., and the mixture was stirred for 5h. After this time, the reaction was filtered and the solvents wereevaporated under reduced pressure. Purification by preparative TLC(silica gel, 30% EtOAc/cyclohexane) afforded 2-153 (13.8 mg, 85%) as amixture of two diastereoisomers. ¹H NMR (400 MHz, CDCl₃, 25° C.):δ=12.33 (s, 1H), 12.11 (s, 1H), 9.45 (s, 1H), 9.40 (s, 1H), 6.67, (m,2H), 6.28 (2×s, 2H), 5.83 (d, J=13.2 Hz, 1H), 5.79 (d, J=12.9 Hz, 1H),5.35-5.30 (m, 3H), 5.27-5.22 (m, 3H), 5.06 (bd, J=8.2 Hz, 2H), 4.10 (d,J=17.5 Hz, 2H), 3.90-3.85 (m, 1H), 3.80-3.76 (m, 1H), 3.65 (d, J=17.7Hz, 2H), 3.57-3.52 (m, 2H), 3.46-3.41 (m, 2H), 2.77-2.71 (m, 3H),2.53-2.49 (m, 3H), 2.36-2.29 (m, 4H), 2.24-1.56 (m, 12H), 1.31 (d, J=6.4Hz, 3H), 1.28 (d, J=6.4 Hz, 3H); HRMS (ESI-TOF): m/z: calculated forC₂₃H₂₈O₆Na: 423.1778, found 423.1778 [M+Na⁺].

General procedure for the synthesis of compounds 2-154: Thehydroxylamine R²ONH₂ (5.0 equiv.) was added to a solution of compound2-120 (1.0 equiv.) in pyridine/AcOH (5:1, 0.03 M) and the mixture washeated up to 40° C. After stirring overnight, the solvents wereevaporated under reduced pressure with silica gel. Purification over ashort pad of silica gel with a mixture of 30% EtOAc/cyclohexane affordedcompound 2-154 (˜99%) as a mixture of two diastereoisomers cis/trans.

Selected example of compounds 2-154: ¹H NMR (400 MHz, CDCl₃, 25° C.):δ=7.50-7.25 (m, 10H), 6.82 (s, 1H), 6.75 (s, 1H), 6.66 (s, 1H), 6.48 (s,1H), 6.24-6.11 (m, 2H), 6.11-6.05 (m, 2H), 5.45-5.38 (m, 4H), 5.34-5.31(m, 14H), 4.50 (d, J=17.2 Hz, 1H), 3.65-3.38 (m, 8H), 3.60 (d, J=17.1Hz, 1H), 3.54 (d, J=17.1 Hz, 1H), 3.24 (d, J=17.2 Hz, 1H), 2.48-2.36 (m,4H), 2.21-2.17 (m, 2H), 2.11-2.04 (m, 2H), 1.95-1.83 (m, 2H), 1.62-1.51(m, 2H), 1.49 (d, J=6.4 Hz, 6H), 1.32-1.20 (m, 12H); ¹³C NMR (100 MHz,CDCl₃, 25° C.): δ=168.1, 167.9, 159.1, 158.8, 157.2, 155.6, 155.4,154.2, 140.8, 138.2, 138.2, 137.8, 136.9, 136.7, 132.3, 132.3, 128.3(×2), 128.3 (×2), 128.2, 128.1 (×2), 128.0 (×2), 127.7, 127.6, 125.5,118.8, 118.6, 118.3, 108.8, 108.5, 101.7, 101.7, 93.5, 93.4, 93.1 (×2),77.2, 76.0, 75.9, 71.2, 71.0, 64.5, 64.5, 64.3, 64.3, 40.0, 40.0, 34.9,32.4, 32.3, 31.6, 31.1, 28.9, 20.3, 20.2, 15.0 (×2), 15.0 (×2); HRMS(ESI-TOF): m/z: calculated for C₃₁H₃₉NO₇Na: 560.2619, found 560.2627[M+Na⁺].

General procedure for the synthesis of compounds 2-155: PS-TsOH (10.0equiv., 3.2 mmol.g⁻¹) was added to a solution of compound 2-154 (1.0equiv.) in MeOH (0.02 M) and the suspension was shaken at 40° C. for 4h. After this time, the reaction mixture was filtered and the methanolicsolution concentrated under reduced pressure. The crude product obtainedwas submitted without further purification to the next step. Thus, DHP(1.0 equiv.) and PS-TsOH (catalytic amount, 3.2 mmol.g⁻¹) were added toa solution of this crude in CH₂Cl₂ (0.02 M) at 23° C., and the mixturewas stirred for 5 h. After this time, the mixture was filtered, thesolvents were evaporated under reduced pressure, and the remainingresidue was purified by preparative TLC (silica gel, 30%EtOAc/cyclohexane) to afford two different diastereoisomers 1:1 of 2-155(˜65% yield).

Selected example of compounds 2-155: Less polar diastereoisomer: ¹H NMR(400 MHz, CDCl₃, 25° C.): δ=9.25 (s, 1H), 9.24 (s, 1H), 7.46-7.33 (m,10H), 6.29 (s, 1H), 6.26 (s, 1H), 6.07-6.02 (m, 2H), 5.75 (d, J=15.8 Hz,1H), 5.69 (d, J=15.8 Hz, 1H), 5.44-5.38 (m, 6H), 5.23 (s, 4H), 5.03 (d,J=8.8 Hz, 2H), 4.34-4.13 (m, 6H), 3.69-3.63 (m, 2H), 2.70-2.67 (m, 2H),2.30-2.16 (m, 6H), 2.08-1.94 (m, 8H), 1.73-1.65 (m, 8H), 1.42 (t, J=6.4Hz, 3H), 1.39 (t, J=7.0 Hz, 3H); HRMS (ESI-TOF): m/z: calculated forC₃₀H₃₅NO₆Na: 528.2357, found 528.2562 [M+Na⁺].

More polar diastereoisomer: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=11.61 (s,1H), 9.27 (s, 1H), 7.41-7.33 (m, 5H), 6.62 (d, J=16.4 Hz, 1H), 6.47 (s,1H), 6.15-6.07 (m, 1H), 5.50-5.38 (m, 3H), 5.16 (s, 2H), 5.04 (d, J=10.5Hz, 1H), 4.30 (d, J=15.2 Hz, 1H), 4.24 (d, J=10.5 Hz, 1H), 3.84 (d,J=15.2 Hz, 1H), 3.66 (t, J=11.4 Hz, 1H), 2.71-2.65 (m, 1H), 2.28-2.08(m, 6H), 1.73-1.64 (m, 5H), 1.38 (t, J=7.0 Hz, 3H); HRMS (ESI-TOF): m/z:calculated for C₃₀H₃₅NO₆Na: 528.2357, found 528.2494 [M+Na⁺].

Macrocycle 2-128 from pochonin D (2-85): TBSCl (53.6 mg, 356 mmol) andimidazole (23.6 mg, 356 μmol) were added to a solution of pochonin D(2-85, 25 mg, 71.2 mmol) in DMF (5 mL) and the mixture was stirred for 3h at room temperature. Purification by column chromatography (silicagel, 0-30% EtOAc/cyclohexane gradient) afforded compound 2-128 (40 mg,98%).

General procedure for compounds 2-157: The hydroxylamine RONH₂ (5.0equiv.) was added to a solution of compound 2-128 (1.0 equiv.) inpyridine/AcOH (5:1, 250 μL) and the mixture was heated up to 40° C.After stirring overnight, the solvents were evaporated under reducedpressure, and filtration on silica gel with a mixture of 30%EtOAc/cyclohexane afforded two isomers of 2-157 (˜90% yield).

Selected example of compounds 2-157: cis oxime: ¹H NMR (400 MHz, CDCl₃,25° C.): δ=7.42 (bd, J=6.4 Hz, 2H), 7.36 (bdd, J=7.5, 6.9 Hz, 2H),7.34-7.32 (m, 1H), 6.52 (d, J=16.1 Hz, 1H), 6.38 (s, 1H), 6.18-6.10 (m,1H), 5.36-5.32 (m, 2H), 5.16 (bs, 2H), 4.99-4.95 (m, 1H), 3.79-3.76 (m,2H), 2.40-1.99 (m, 6H), 1.45 (d, J=6.2 Hz, 3H), 1.03 (s, 9H), 0.99 (s,9H), 0.28 (s, 3H), 0.26 (s, 3H), 0.20 (s, 6H); trans oxime: ¹H NMR (400MHz, CDCl₃, 25° C.): δ=7.44 (bd, J=6.5 Hz, 2H), 7.37 (bdd, J=7.6, 6.9Hz, 2H), 7.33-7.31 (m, 1H), 6.41 (s, 1H), 6.04-5.97 (m, 1H), 5.48 (bd,J=15.0 Hz, 1H), 5.29-5.27 (m, 1H), 5.22 (bs, 2H), 5.00-4.95 (m, 1H),3.98-3.89 (m, 2H), 2.39-2.02 (m, 6H), 1.37 (d, J=5.9 Hz, 3H), 1.04 (s,9H), 0.99 (s, 9H), 0.28 (s, 3H), 0.27 (s, 3H), 0.23 (s, 3H), 0.22 (s,3H).

General procedure for compounds 2-158: To a solution of correspondingcompound 2-157 (1.0 equiv) in THF was added TBAF (2.5 equiv, 1M solutionin THF) and the mixture was stirred at room temperature for 2 h. Thesolvents were then evaporated under reduced pressure, and filtration onsilica gel with a mixture of 30% EtOAc/cyclohexane afforded compound2-158 in >85% yield.

Selected example of compounds 2-158: cis oxime: ¹H NMR (400 MHz, CDCl₃,25° C.): δ=11.52 (s, 1H), 7.45-7.34 (m, 5H), 6.64 (s, 1H), 6.09-6.02 (m,2H), 5.34-5.25 (m, 4H), 5.18-5.08 (m, 2H), 4.33 (d, J=17.0 Hz, 1H), 4.15(d, J=17.6 Hz, 1H), 2.65-2.59 (m, 1H), 2.27-2.14 (m, 3H), 2.04-2.00 (m,1H), 1.88-1.83 (m, 1H), 1.30 (t, J=6.4 Hz, 3H); HRMS (ESI-TOF): m/z:calculated for C₂₅H₂₆ClNO₅Na: 478.1392, found 478.1372 [M+Na⁺].

trans oxime: ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=11.73 (s, 1H), 7.32-7.26(m, 5H), 6.64 (s, 1H), 6.50 (d, J=16.4 Hz, 1H), 6.06-5.98 (m, 2H),5.43-5.24 (m, 3H), 4.91 (s, 2H), 4.22 (s, 2H), 2.61-2.55 (m, 1H),2.46-2.33 (m, 2H), 2.20-2.02 (m, 3H), 0.98 (t, J=6.4 Hz, 3H); HRMS(ESI-TOF): m/z: calculated for C₂₅H₂₆ClNO₅Na: 478.1392, found 478.1522[M+Na⁺].

Compound (±)-2-159: A solution of cis-butene oxide (1.75 mL, 20 mmol) inEt₂O (10 mL) was cooled to −30° C. Copper iodide (1.14 g, 6 mmol) wasadded to this solution and then, vinyl magnesium bromide (40 mL, 1Msolution in THF, 40 mmol) was added dropwise over a period of 1 h. Thereaction mixture was then warmed up to room temperature over 12 h andthe reaction turned black. The reaction mixture was quenched slowly withsaturated NH₄Cl_(aq.) (20 mL), stirred for 2 h, extracted with Et₂O (20mL) and dried over MgSO₄. Concentration under reduced pressure affordedcompound (+)-2-159 (1.3 g, 65%). ¹H NMR (400 MHz, CDCl₃, 25° C.):δ=5.81-5.72 (m, 1H), 5.15 (d, J=13.2 Hz, 2H), 3.59 (m, 1H), 2.23-2.10(m, 1H), 1.21 (d, J=6.4 Hz, 3H), 1.05 (d, J=7.0 Hz, 3H).

Compound (±)-2-163: In a similar manner as that described for compound2-112, compound (±)-2-163 was prepared with a 57% yield from (±)-2-162.¹H NMR (400 MHz, CDCl₃, 25° C.): δ=7.10 (s, 1H), 6.76-6.71 (m, 1H), 5.87(d, J=15.8 Hz, 1H), 5.32 (s, 2H), 5.25-5.18 (s+m, 4H), 4.87-4.80 (m,1H), 3.99 (d, J=16.9 Hz, 1H), 3.80-3.71 (m, 5H), 2.32-2.26 (m, 2H),2.20-2.11 (m, 3H), 1.36 (d, J=6.4 Hz, 3H), 1.25 (t, J=7.0 Hz, 6H), 1.04(d, J=7.0 Hz, 3H); ¹³C NMR (100 MHz, CDCl₃, 25° C.): δ=195.7, 166.6,154.6, 153.8, 147.1, 133.3, 132.8, 120.7, 117.7, 102.9, 93.9, 93.6,76.2, 64.8, 64.6, 45.1, 41.6, 30.7, 30.7, 18.2, 16.9, 15.0, 15.0.

Compound (±)-2-164: In a similar manner as that described for compound2-85 using PS-TsOH, compound (±)-2-164 was prepared with a 40% yieldfrom (±)-2-163. ¹H NMR (400 MHz, CDCl₃, 25° C.): δ=11.52 (s, 1H),6.78-6.69 (m, 1H), 6.67 (s, 1H), 6.05 (s, 1H), 5.93 (d, J=16.4 Hz, 1H),5.47-5.45 (m, 1H), 5.37-5.33 (m, 1H), 5.16 (dd, J=15.8, 7.0 Hz, 1H),4.48 (bs, 1H), 4.35 (d, J=17.6 Hz, 1H), 2.45-2.26 (m, 5H), 1.27 (d,J=6.4 Hz, 3H), 1.10 (d, J=7.0 Hz, 3H); ESI: m/z: calculated forC₁₈H₂₁ClO₅: 365.12, found 365.22 [M+H⁺].

The description and examples provided herein are merely illustrative,and the invention is not so limited. Numerous variations, permutationsand derivatives of these compounds, procedures and uses will occur tothose of ordinary skill in the art, and are contemplated within thescope of the invention.

1) A compound of formula I, tautomers thereof, or a pharmaceuticallyacceptable salt, solvate, ester or prodrug thereof:

wherein: R¹, R², R³, R⁴ and R⁵ are independently hydrogen, halogen,nitro, cyano, alkyl, lower alkyl, substituted alkyl, alkenyl, alkynyl,alkylaryl, aralkyl, aryl, heteroalkyl, alkylheteroaryl, heterocyclyl,heteroaryl, OH, OR, NH₂, N(R)₂, SR, S(O)R, S(O)₂R, —SO₂N(R)₂, —N(R)SO₂R,—N(CO)R, —N(CO)N(R)₂, —N(CO)OR, —O(CO)R, —(CO)R, —(CO)OR, —(CO)N(R)₂,—O(CO)OR, or —O(CO)N(R)₂, wherein each R can be the same or different;A₁ and A₂ together are —CH₂—CH₂—, —CH═CH—, —CH(OH)—CH(OH)—,—CH(OH)—CH(halogen)-, —CH(halogen)-CH(OH)—, 1,2-cyclopropadiyl, or1,2-oxirane; B₁ and B₂ together are —CH₂—CH₂— or B₁ and B₂ togetherrepresent a covalent bond; X¹ is hydrogen, halogen, OH, OR, NH₂, N(R)₂,NH—OR, SR, S(O)R, S(O)₂R, —NH—O—(CH₂)_(n)—CO₂—R,—NH—O—(CH₂)_(n)—CON(R)₂; or X₁ together with X₂ or X₃ represents acovalent bond, wherein each R can be the same or different; X² and X³are both hydrogen, or one of X₂ and X₃ is hydrogen and the othertogether with X₁ represents a covalent bond; X⁴ and X⁵ together are ═O,═S, ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂, ═N—N(R)₂, ═N—N—SORor ═N—N—SO₂R; or one of X⁴ and X⁵ is hydrogen and the other is OH, OR,O(CO)R, O(CO)OR, O(CO)N(R)₂, —(CH₂)_(n)C(O)OR, or —(CH₂)_(n)C(O)N(R)₂,and n is 0, 1, 2, or 3; or one of X⁴ and X⁵ together with X⁶ representsa covalent bond and the other of X⁴ and X⁵ is OH, OR, O(CO)R, O(CO)OR,—N(R)₂ or O(CO)N(R)₂, wherein each R can be the same or different; X⁶ ishydrogen or X⁶ together with one of X⁴ and X⁵ represents a covalentbond; and R is hydrogen, alkyl, lower alkyl, acyl, aryl, alkaryl,arylalkyl including benzyl, heteroalkyl, heteroaryl, heterocyclyl, aprotecting group; or two R on the same nitrogen are taken together withthe nitrogen to form a 5-8 membered heterocyclic or heteroaryl ring; andn is 0, 1, 2 or
 3. 2) The compound of claim 1, wherein the compound hasthe structure of formula II:

wherein, R⁷ is ═O, ═S, ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂,═N—N(R)₂, ═N—N—SOR or ═N—N—SO₂R; and wherein each R can be the same ordifferent. 3) The compound of claim 2, wherein R¹ is H, halogen orheterocyclyl. 4) The compound of claim 2, wherein R⁵ is hydrogen, alkyl,lower alkyl, aryl, heteroaryl or arylalkyl. 5) The compound of claim 2,wherein A₁ and A₂ together are —CH═CH—. 6) The compound of claim 2,wherein A₁ and A₂ together are —CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or—CH(halogen)-CH(OH)—. 7) The compound of claim 2, wherein A₁ and A₂together are 1,2-oxirane. 8) The compound of claim 2, wherein: R¹ is H,Cl or heterocyclyl; R² and R⁴ are independently OH or OR; R⁵ ishydrogen, alkyl, aryl or aralkyl; A₁ and A₂ together are —CH═CH— or—C(OH)—C(OH)—; X¹ is hydrogen, halogen or NH—OR; and R⁷ is ═O, ═S,═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂, ═N—N(R)₂, ═N—N—SOR,═N—N—SO₂R. 9) The compound of claim 2, wherein: R¹ is H, Cl orheterocyclyl; R² and R⁴ are independently OH or OR; R⁵ is hydrogen,alkyl, aryl or aralkyl; A₁ and A₂ together are 1,2-oxirane; X¹ ishydrogen, halogen or NH—OR; and R⁷ is ═O, ═S, ═N—OR, ═N—O—(CH₂)_(n)COOR,═N—O—(CH₂)_(n)CON(R)₂, ═N—N(R)₂, ═N—N—SOR, ═N—N—SO₂R. 10) The compoundof claim 2, wherein: R¹ is H, Cl or heterocyclyl; R² and R⁴ areindependently OH or OR; R⁵ is hydrogen, alkyl, aryl or aralkyl; A₁ andA₂ together are —CH═CH— or —C(OH)—C(OH)—; X¹ together with X² representa bond; and R⁷ is ═O, ═S, ═N—OR, ═N—O—(CH₂)_(n)COOR,═N—O—(CH₂)_(n)CON(R)₂, ═N—N(R)₂, ═N—N—SOR, ═N—N—SO₂R. 11) The compoundof claim 2, wherein: R¹ is H, Cl or heterocyclyl; R² and R⁴ areindependently OH or OR; R⁵ is hydrogen, alkyl, aryl or aralkyl; A₁ andA₂ together are 1,2-oxirane; X¹ together with X² represent a bond; andR⁷ is ═O, ═S, ═N—OR, ═N—O—(CH₂)_(n)COOR, ═N—O—(CH₂)_(n)CON(R)₂,═N—N(R)₂, ═N—N—SOR, ═N—N—SO₂R. 12) The compound of claim 10, wherein: R¹is H or Cl; R⁵ is hydrogen, methyl, propyl, isopropyl or phenyl; and R⁷is ═N—OR, ═N—O—(CH₂)_(n)COOR, or ═N—O—(CH₂)_(n)CON(R)₂. 13) The compoundof claim 12, wherein R¹ is Cl and R⁵ is hydrogen. 14) The compound ofclaim 12, wherein R⁷ is ═N—O—(CH₂)_(n)COOR, or ═N—O—(CH₂)_(n)CON(R)₂;and n is
 1. 15) The compound of claim 12, wherein R⁵ is hydrogen and R⁷is ═N—O—(CH₂)_(n)COOR, or ═N—O—(CH₂)_(n)CON(R)₂. 16) The compound ofclaim 12, wherein R⁵ is hydrogen and R⁷ is ═N—OR. 17) The compound ofclaim 8, wherein R₇ is ═O. 18) The compound of claim 8, wherein R₇ is═N—OR, ═N—O—(CH₂)_(n)COOR, or ═N—O—(CH₂)_(n)CON(R)₂. 19) The compound ofclaim 11, wherein R₇ is ═O. 20) The compound of claim 11, wherein R₇ is═N—OR, ═N—O—(CH₂)_(n)COOR, or ═N—O—(CH₂)_(n)CON(R)₂. 21) The compound ofclaim 1, wherein the compound has the formula III:

wherein R is hydrogen, alkyl, arylalkyl, acyl or a protecting group. 22)The compound of claim 21, wherein R is hydrogen or acyl; and R₁ is H,halogen or heterocyclyl. 23) The compound of claim 21, wherein R⁵ ishydrogen, alkyl, lower alkyl, aryl, heteroaryl or arylalkyl. 24) Thecompound of claim 21, wherein X¹ together with X₂ represent a covalentbond. 25) The compound of claim 21, wherein X¹ is hydrogen, halogen,NH—OR NH—O—(CH₂)_(n)COOR, or NH—O—(CH₂)_(n)CON(R)₂. 26) The compound ofclaim 21, wherein A₁ and A₂ together are —CH═CH—. 27) The compound ofclaim 21, wherein A₁ and A₂ together are —CH(OH)—CH(OH)—,—CH(OH)—CH(halogen)- or —CH(halogen)-CH(OH)—. 28) The compound of claim21, wherein A₁ and A₂ together are 1,2-oxirane. 29) The compound ofclaim 1, wherein the compound has the formula IV:

wherein R⁶ is hydrogen, OR, or N(R)₂. 30) The compound of claim 29,wherein R is hydrogen or acyl. 31) The compound of claim 29, wherein R¹is H, halogen or heterocyclyl. 32) The compound of claim 29, wherein R⁵is hydrogen, alkyl, lower alkyl, aryl, heteroaryl or arylalkyl. 33) Thecompound of claim 29, wherein A₁ and A₂ together are —CH═CH—. 34) Thecompound of claim 29, wherein A₁ and A₂ together are —CH(OH)—CH(OH)—,—CH(OH)—CH(halogen)- or —CH(halogen)-CH(OH)—. 35) The compound of claim29, wherein A₁ and A₂ together are 1,2-oxirane. 36) The compound ofclaim 1, wherein the compound has the formula V:

wherein R⁶ is (CH₂)_(n)C(O)OR, or —(CH₂)_(n)C(O)N(R)₂; and n is 0, 1, 2or
 3. 37) The compound of claim 36, wherein R⁶ is —CH₂C(O)N(Me)OMe. 38)The compound of claim 36, wherein R¹ is H, halogen or heterocyclyl. 39)The compound of claim 36, wherein R⁵ is hydrogen, alkyl, lower alkyl,aryl, heteroaryl or arylalkyl. 40) The compound of claim 36, wherein A₁and A₂ together are —CH═CH—. 41) The compound of claim 36, wherein A₁and A₂ together are —CH(OH)—CH(OH)—, —CH(OH)—CH(halogen)- or—CH(halogen)-CH(OH)—. 42) The compound of claim 36, wherein A₁ and A₂together are 1,2-oxirane. 43) The compound of claim 1, wherein thecompound is selected from the group consisting of

44) A pharmaceutical composition comprising an effectivekinase-inhibiting amount of a compound of any of claim 1, 2, 21, 29, 36or 43 in combination with a pharmaceutically acceptable carrier. 45) Apharmaceutical composition comprising an effective HSP 90-inhibitingamount of a compound of any of claim 1, 2, 21, 29, 36 or 43 incombination with a pharmaceutically acceptable carrier. 46) Thecomposition of claim 44 or 45, wherein the composition comprisesparticles that are less than about 2 microns average particle size. 47)The composition of claim 44 or 45, wherein the composition isincorporated into a biodegradable or non-biodegradable polymer. 48) Thecomposition of claim 44 or 45, further comprising an additive. 49) Thecomposition of claim 48, wherein the additive is selected from ananti-oxidant, a buffer, a bacteriostat, a liquid carrier, a solute, asuspending agent, a thickening agent, a flavoring agent, a gelatin,glycerin, a binder, a lubricant, an inert diluent, a preservative, asurface active agent, a dispersing agent, a biodegradable polymer, orany combination thereof. 50) The composition of claim 44 or 45, whereinthe carrier is suitable for oral, parenteral, inhalation, topical, orintradermal administration. 51) A method of treating a patient with adisease comprising administering to the patient with the disease aneffective amount of a compound of claim 1, wherein the disease is anautoimmune disease, inflammatory disease, neurological orneurodegenerative disease, cancer, cardiovascular disease, allergy,asthma, or a hormone-related disease. 52) A method of treating a patientwith a cancer comprising administering to the patient having the canceran effective cancer-treating amount of a compound of claim
 1. 53) Themethod of claim 52, wherein the cancer is a solid tumor, blood bornetumor, breast, ovary, cervix, prostate, testis, genitourinary tract,esophagus, larynx, glioblastoma, stomach, skin, keratoacanthoma, lung,epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lungadenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma, thyroid,follicular carcinoma, undifferentiated carcinoma, papillary carcinoma,seminoma, melanoma, sarcoma, bladder carcinoma, liver carcinoma andbiliary passages, kidney carcinoma, myeloid disorders, lymphoiddisorders, Hodgkin's, hairy cells, buccal cavity, pharynx, lip, tongue,mouth, pharynx, small intestine, colon-rectum, large intestine, rectum,brain and central nervous system, or leukemia. 54) A method of treatinga patient with a disease associated with undesirable neovascularizationcomprising administering to the patient with the undersirableneovascularization an effective amount of a compound of claim
 1. 55) Themethod of claim 54, wherein the disease associated with undesirableneovasculariation comprises ocular neovascular disease, diabeticretinopathy, retinopathy of prematurity, corneal graft rejection,neovascular glaucoma and retrolental fibroplasias, epidemickeratoconjunctivitis, Vitamin A deficiency, contact lens overwear,atopic keratitis, superior limbic keratitis, pterygium keratitis sicca,Sjögren's syndrome, acne rosacea, phylectenulosis, syphilis,Mycobacteria infections, lipid degeneration, chemical burns, bacterialulcers, fungal ulcers, Herpes simplex infections, Herpes zosterinfections, protozoan infections, Kaposi's sarcoma, Mooren's ulcer,Terrien's marginal degeneration, marginal keratolysis, trauma,rheumatoid arthritis, systemic lupus, polyarteritis, Wegener'ssarcoidosis, Scleritis, Steven-Johnson disease, pemphigoid, radialkeratotomy, or corneal graph rejection, sickle cell anemia, sarcoid,pseudoxanthoma elasticum, Paget's disease, vein occlusion, arteryocclusion, carotid obstructive disease, chronic uveitis/vitritis, Lyme'sdisease, systemic lupus erythematosis, Eales' disease, Bechet's disease,infections causing a retinitis or choroiditis, presumed ocularhistoplasmosis, Best's disease, myopia, optic pits, Stargart's disease,pars planitis, chronic retinal detachment, hyperviscosity syndromes,toxoplasmosis, or post-laser complications. 56) A method of treating apatient with an inflammatory disease associated with inflammationcomprising administering to the patient with the inflammatory disease aneffective amount of a compound of claim
 1. 57) The method of claim 56,wherein the inflammatory disease is excessive or abnormal stimulation ofendothelial cells, atherosclerosis, vascular malfunctions, abnormalwound healing, inflammatory and immune disorders, Bechet's disease, goutor gouty arthritis, abnormal angiogenesis accompanying rheumatoidarthritis, skin diseases, psoriasis, diabetic retinopathy, retinopathyof prematurity, retrolental fibroplasia, macular degeneration, cornealgraft rejection, neovascular glaucoma or Osler Weber syndrome.